US10308596B2 - HDAC8 inhibitors for treating cancer - Google Patents

HDAC8 inhibitors for treating cancer Download PDF

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US10308596B2
US10308596B2 US15/042,012 US201615042012A US10308596B2 US 10308596 B2 US10308596 B2 US 10308596B2 US 201615042012 A US201615042012 A US 201615042012A US 10308596 B2 US10308596 B2 US 10308596B2
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US20160237027A1 (en
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Ya-Huei Kuo
Wei-Jan Huang
Chung-I Chang
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Academia Sinica
Taipei Medical University TMU
City of Hope
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Taipei Medical University TMU
City of Hope
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    • C07C259/06—Compounds containing carboxyl groups, an oxygen atom of a carboxyl group being replaced by a nitrogen atom, this nitrogen atom being further bound to an oxygen atom and not being part of nitro or nitroso groups without replacement of the other oxygen atom of the carboxyl group, e.g. hydroxamic acids having carbon atoms of hydroxamic groups bound to hydrogen atoms or to acyclic carbon atoms
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    • A61P35/00—Antineoplastic agents
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    • C07D209/04—Indoles; Hydrogenated indoles
    • C07D209/08—Indoles; Hydrogenated indoles with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to carbon atoms of the hetero ring
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    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/04—Indoles; Hydrogenated indoles
    • C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
    • C07D209/18—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D213/54—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
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    • C07D215/12—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
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    • C07D217/00—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems
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    • C07D217/14—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with radicals, substituted by hetero atoms, attached to carbon atoms of the nitrogen-containing ring other than aralkyl radicals
    • C07D217/16—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with radicals, substituted by hetero atoms, attached to carbon atoms of the nitrogen-containing ring other than aralkyl radicals substituted by oxygen atoms
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    • C07D217/18—Aralkyl radicals
    • C07D217/20—Aralkyl radicals with oxygen atoms directly attached to the aromatic ring of said aralkyl radical, e.g. papaverine
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    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D307/91—Dibenzofurans; Hydrogenated dibenzofurans
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    • C07D317/00—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms
    • C07D317/08—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3
    • C07D317/44—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D317/46—Heterocyclic compounds containing five-membered rings having two oxygen atoms as the only ring hetero atoms having the hetero atoms in positions 1 and 3 ortho- or peri-condensed with carbocyclic rings or ring systems condensed with one six-membered ring
    • C07D317/48—Methylenedioxybenzenes or hydrogenated methylenedioxybenzenes, unsubstituted on the hetero ring
    • C07D317/50—Methylenedioxybenzenes or hydrogenated methylenedioxybenzenes, unsubstituted on the hetero ring with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to atoms of the carbocyclic ring
    • C07D317/60—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C—CHEMISTRY; METALLURGY
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    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
    • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
    • C07D333/06—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to the ring carbon atoms
    • C07D333/24—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals

Definitions

  • Histone deacetylases play a role in the reversible acetylation of histones, transcription factors, and other proteins, which are associated with chromatin remodeling and regulation of gene expression.
  • AML Acute myeloid leukemia
  • AML arises, in part, from disordered hematopoiesis as a consequence of multiple cooperative mutations or alternations disrupting differentiation, proliferation and survival programs in hematopoietic progenitors.
  • Recurrent chromosomal abnormalities in AML frequently involve transcription factor fusion proteins that contribute to unique etiology and prognosis (1).
  • HDAC8 activity is linked to cancer.
  • compounds and methods for treating cancer with HDAC8 inhibitors are provided herein.
  • A is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.
  • X is —C(R 4 ) ⁇ or —N ⁇ .
  • Y is a bond, —N(R 5 )—, —O—, or —S—.
  • L 1 is a bond, —C(O)—, —C(O)O—, —O—, —S—, —N(R 6 )—, —C(O)N(R 6 )—, —S(O) n6 —, —S(O)N(R 6 )—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene.
  • R 1 is halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 1A , —C(O)R 1A , —NR 1A R 1B , —C(O)OR 1A , —C(O)NR 1A R 1B , —NO 2 , —SR 1A , —S(O) n1 R 1A , —S(O) n1 OR 1A , —S(O) n1 NR 1A R 1B , —NHNR 1A R 1B , —ONR 1A R 1B , —NHC(O)NHNR 1A R 1B , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substitute
  • R 2 is halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 2A , —C(O)R 2A , —NR 2A R 2B , —C(O)OR 2A , —C(O)NR 2A R 2B , —NO 2 , —SR 2A , —S(O) n2 R 2A , —S(O) n2 OR 2A , —S(O) n2 NR 2A R 2B , —NHNR 2A R 2B , —ONR 2A R 2B , —NHC(O)NHNR 2A R 2B , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 3 is independently halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —S, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 4 is hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 4A , —C(O)R 4A , —NR 4A R 4B , —C(O)OR 4A , —C(O)NR 4A R 4B , —NO 2 , —SR 4A , —S(O) n4 R 4A , —S(O) n4 OR 4A , —S(O) n4 NR 4A R 4B , —NHNR 4A R 4B , —ONR 4A R 4B , —NHC(O)NHNR 4A R 4B , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 5 is hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 5A , —C(O)R 5A , —NR 5A R 5B , —C(O)OR 5A , —C(O)NR 5A R 5B , —NO 2 , —SR 5A , —S(O) n5 R 5A , —S(O) n5 OR 5A , —S(O) n5 NR 5A R 5B , —NHNR 5A R 5B , —ONR 5A R 5B , —NHC(O)NHNR 5A R 5B , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 6 is hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 6A , —C(O)R 6A , —NR 6A R 6B , —C(O)OR 6A , —C(O)NR 6A R 6B , —NO 2 , —SR 6A , —S(O) n6 R 6A , —S(O) n6 OR 6A , —S(O) n6 NR 6A R 6B , —NHNR 6A R 6B , —ONR 6A R 6B , —NHC(O)NHNR 6A R 6B , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl
  • R 1A , R 1B , R 2A , R 2B , R 4A , R 4B , R 5A , R 5B , R 6A , and R 6B are independently hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 Cl, —S(O) 3 H, —S(O) 4 H, —S(O) 2 NH 2 , —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , —NHC(O)NH 2 , —NHS(O) 2 H, —NHC(O)H, —NHC(O)—OH, —NHOH, —OCF 3 , —OCHF 2 , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl
  • HDAC8 mediated deacetylation of p53 methods of inhibiting HDAC8 mediated deacetylation of p53.
  • a method of inhibiting HDAC8 mediated deacetylation of p53 by contacting HDAC8 with a HDAC8 inhibitor in the presence of p53, thereby inhibiting HDAC8 deacetylation of p53.
  • FIGS. 1A-1G CBF ⁇ -SMMHC expression impaired p53 target gene induction and reduced acetylation of p53.
  • FIG. 1A Relative expression of p53 target genes including (in order left to right, top to bottom) p21, Mdm2, Bax, Bid, Puma, Gadd45b, LincRNA-p21 and Stag1 in 32D-CM or 32D-CBF ⁇ cells, determined by qRT-PCR. Shown are fold induction 24 h after 3 Gy IR relative to non-IR (mean+/ ⁇ SD) performed in replicate and three independent experiments.
  • FIG. 1A Relative expression of p53 target genes including (in order left to right, top to bottom) p21, Mdm2, Bax, Bid, Puma, Gadd45b, LincRNA-p21 and Stag1 in 32D-CM or 32D-CBF ⁇ cells, determined by qRT-PCR. Shown are fold induction 24 h after 3 Gy IR relative to non-
  • FIG. 1B Western blot analysis of Ac-p53, p53, CM, CBF ⁇ and HDAC8 at indicated time points (2, 4, 6 and 12 h) after IR in 32D-CM or 32D-CBF ⁇ cells. ⁇ -actin served as loading control.
  • FIG. 1C Western blot time course analysis (2, 4, 6 and 12 h) of Ac-p53, p53, CM, CBF ⁇ after IR in BM progenitor cells isolated from CM-expressing pre-leukemic or control mice. ⁇ -actin served as loading control.
  • FIG. 1B Western blot analysis of Ac-p53, p53, CM, CBF ⁇ and HDAC8 at indicated time points (2, 4, 6 and 12 h) after IR in 32D-CM or 32D-CBF ⁇ cells. ⁇ -actin served as loading control.
  • FIG. 1C Western blot time course analysis (2, 4, 6 and 12 h) of Ac-p53, p53, CM, CBF ⁇ after
  • FIG. 1D Western blot of Ac-p53, p53, CM, CBF ⁇ in Cbfb 56M/+ BM progenitor cells transduced with MIG-Cre and IR (3 Gy, 6 h). ⁇ -actin served as loading control.
  • FIG. 1E Western blot of Ac-p53, p53, CM, CBF ⁇ in 32D-CM cells expressing control (Ctrl)-shRNA or CM shRNA (A3, D4) 6 h after 3 Gy IR. ⁇ -actin served as loading control.
  • FIG. 1E Western blot of Ac-p53, p53, CM, CBF ⁇ in 32D-CM cells expressing control (Ctrl)-shRNA or CM shRNA (A3, D4) 6 h after 3 Gy IR. ⁇ -actin served as loading control.
  • FIG. 1F Western blot of CM and ⁇ -actin in sorted leukemic BM cells transduced with non-silencing ctrl-shRNA or CM shRNA (A3, D4).
  • FIG. 1G Histogram depicting relative expression of p53 target genes including (in order left to right) p21, Gadd45b, LincRNA-p21 Bax, Puma, and Bid in sorted leukemic BM cells transduced with non-silencing ctrl-shRNA or CM shRNA (A3, D4). Shown are fold change (mean+/ ⁇ SD) relative to ctrl-shRNA expressing cells, performed in triplicate. Ordering (left to right for p53 target): ctrl, A3, A4.
  • FIGS. 2A-2F CBF ⁇ -SMMHC fusion protein aberrantly interacts with p53.
  • FIG. 2A Co-IP and immunoblot (IB) analysis in 32D-CM (top) or 32D-CBF ⁇ (bottom) cells using anti-p53 or anti-mouse IgG for IP, and anti-p53 (left) or anti-CBF ⁇ (right) for IB. Shown are representative of more than three independent experiments.
  • FIG. 2B Co-IP (anti-FLAG) and IB (anti-p53) analysis in 32D-Flag, 32D-CBF ⁇ or 32D-CM cells. Input of CBF ⁇ or CM are shown using anti-FLAG (IP, IB).
  • FIG. 2C Co-IP and IB analysis in control BM progenitor cells (lane 1 from left), CM-expressing pre-leukemic or leukemic BM cells without IR (left) or pre-leukemic BM cells after IR (right). IP was performed with anti-CBF ⁇ and IB was performed with anti-p53 (top) or anti-CBF ⁇ (middle). Bottom panel shows western blot using anti-p53.
  • FIG. 2D Co-IP (anti-FLAG) and IB (anti-p53) analysis in nuclear and cytoplasmic fractions prepared from the indicated cell lines. Western blot analyses for CM, p53, and Histone H3 for each fraction are shown.
  • FIG. 2E DUOLINK® in situ proximity ligation assay (PLA) using mouse anti-CBF ⁇ antibody plus rabbit anti-p53 antibody and PLA probes. Punctuate red fluorescent spots indicate intermolecular protein interactions (left). DAPI-stained nuclei are shown in blue (center) and GFP reporter indicates transduced cells (right). Shown are representative images.
  • FIG. 2F Co-IP and IB in inv(16) + AML (163, 987) or non-inv(16) AML (467, 865) CD34 + cells 3 h after IR. Co-IP was performed using anti-p53 (DO-1) or anti-mouse IgG, and IB was performed with anti-CBF ⁇ (top) or anti-p53 antibodies (bottom).
  • FIGS. 3A-3F CBF ⁇ -SMMHC recruits p53 and HDAC8 in a protein complex through distinct protein regions.
  • FIG. 3A Sequential co-IP and IB analysis in 32D-CBF ⁇ or 32D-CM cells using anti-HDAC8 for the primary IP, followed by IP using anti-CBF ⁇ , and IB with anti-p53. Cells were either not IR (left panel) or received 3 Gy IR (right panel).
  • FIG. 3B Sequential co-IP and IB analysis in 32D-CBF ⁇ or 32D-CM cells using anti-p53 for the primary IP, followed by IP using anti-CBF ⁇ , and IB with anti-HDAC8.
  • FIG. 3A Sequential co-IP and IB analysis in 32D-CBF ⁇ or 32D-CM cells using anti-p53 for the primary IP, followed by IP using anti-CBF ⁇ , and IB with anti-HDAC8.
  • FIG. 3C Illustration of CM deletion variants (left) used in Co-IP (anti-p53) and IB (anti-CBF ⁇ or anti-p53) analysis (right). Arrows correspond to the expected size of CM variants.
  • FIG. 3D DUOLINK® in situ PLA in 32D cells expressing FL-CM or deletion mutants d134, d179 or DC95 using mouse anti-CBF ⁇ plus rabbit anti-p53 and PLA probes. Red fluorescent spots indicate CM-p53 protein interactions (top), DAPI staining is in blue (center) and GFP reporter indicates transduced cells (bottom). Shown are representative images.
  • FIG. 3D Illustration of CM deletion variants (left) used in Co-IP (anti-p53) and IB (anti-CBF ⁇ or anti-p53) analysis (right). Arrows correspond to the expected size of CM variants.
  • FIG. 3D DUOLINK® in situ PLA in 32D cells expressing FL-CM or deletion mutants d134, d179 or DC95 using mouse
  • FIG. 3E Western blotting of Hdac8 in 32D-CM cells expressing non-silencing control shRNA or Hdac8-shRNA (sh1 or sh2) (left). Co-IP (IgG or anti-p53) and IB (anti-CBF ⁇ or anti-p53) analysis in shRNA (control, sh1 or sh2) expressing cells (right).
  • FIG. 3F In situ PLA in 32D-CM cells expressing control shRNA or Hdac8-shRNA (sh1 or sh2) using mouse anti-CBF ⁇ plus rabbit anti-p53 and PLA probes. Spots indicate CM-p53 protein interactions (left), DAPI staining (center) and GFP reporter indicates transduced cells (right). Representative images are shown.
  • FIGS. 4A-4E HDAC8 mediates the deacetylation of p53 in CBF ⁇ -SMMHC-expressing cells.
  • FIG. 4A Western blotting of Hdac8, Ac-p53 (K379), and total p53 levels after shRNA-mediated knock-down in 32D-CM cells in response to IR (3 Gy). Levels of ⁇ -actin were detected as loading control.
  • FIG. 4B Western blotting of Ac-p53 (K379), p53 and Hdac8 in 32D-CM cells treated with HDAC8 inhibitor PCI-34051, 22d or Nutlin-3 at doses indicated for 6 h.
  • FIG. 4C Western blotting of Ac-p53 (K379), p53 in CM, ⁇ C95 compared to CBF ⁇ or FLAG expressing 32D cells at 2 h or 4 h after IR (3 Gy).
  • FIG. 4D Fold induction of p53 target genes including p21, Mdm2, Bax, Bid, Puma, Gadd45b, LincRNA-p21 and Stag1 in 32D-CBF ⁇ , CM or CM- ⁇ C95 expressing cells, determined by qRT-PCR. Shown are fold induction 24 h after 3 Gy IR relative to non-IR (mean+/ ⁇ SD) performed in triplicate and two independent experiments.
  • FIGS. 5A-5G Pharmacological inhibition of HDAC8 selectively activates p53, reduces proliferation and induces p53-dependent apoptosis in inv(16) + AML CD34 + cells.
  • FIG. 5D Western blotting of Ac-p53 (K382), and p53 levels in inv(16) + AML CD34 + cells treated with 22d (10 ⁇ M) or Nutlin-3 (2.5 ⁇ M) for 6 h. Levels of ⁇ -actin were detected as loading control. Shown are representative results from four patients.
  • FIG. 5F Representative FACS plot of Annexin V/DAPI labeling in inv(16) + AML CD34 + cells transduced with a pLKO-GFP vector expressing p53 shRNA (GFP + ) and treated with 22d for 48 h.
  • FIG. 5G Relative survival of sorted GFP+ inv(16) + AML CD34 + cells expressing p53 shRNA (open diamond) or non-silencing control (solid dot) and treated with indicated doses of 22d for 48 h. Each dot represents an individual patient and lines indicate mean ⁇ SEM.
  • FIGS. 6A-6G Inhibiting HDAC8 by pharmacological inhibitor 22d eliminates LSC engraftment and AML propagation.
  • FIG. 6A Schematic illustration of experimental design. Cbfb +/56M Mx1Cre or Cbfb +/56M Mx1Cre/tdTomato + mice were induced with pIpC and AML cells were isolated from bone marrow of moribund mice, treated with 22d or vehicle for 48 or 72 h and transplanted into sub-lethally irradiated (6.5 Gy) C57Bl/6 congenic mice.
  • FIG. 6C Representative images of spleens from mice transplanted with vehicle treated cells (top) or 22d treated cells (bottom) 8 weeks after transplantation.
  • FIG. 6D Representative FACS plots of engrafted dTomato + /cKit + AML cells in the bone marrow or spleen 8 weeks after transplantation. Shown are representative frequencies of dTomato + /cKit + cells in individual transplanted mice.
  • FIG. 7A-7I In vivo administration of 22d significantly reduce AML burden and abrogate LSC activity.
  • FIG. 7A Schematic illustration of experimental design. Cbfb +/56M Mx1Cre/tdTomato + mice were induced with pIpC and AML cells were isolated from bone marrow of moribund mice and transplanted into sub-lethally irradiated (6.5 Gy) C57Bl/6 congenic mice. After 5-6 weeks, mice were randomized into two groups, one group was treated with vehicle and the other treated with 22d by intraperitoneal injection (50 mg/kg/dose) twice a day for 2 weeks. AML engraftment was analyzed at the end of the treatment period, and transplanted into 2 nd recipients.
  • FIG. 7B Representative FACS plots of engrafted dTomato+/cKit+ AML cells in the bone marrow after the 2-week treatment with vehicle (top) or 22d (bottom).
  • FIG. 7B Representative FACS plots of engrafted dTomato+/cKit+ AML cells in the bone marrow after the 2-week treatment with vehicle (top) or 22d (bottom).
  • FIG. 7E Representative FACS plots of dTomato + /cKit + AML cells in the bone marrow of 2 nd transplant recipients whom received BM from vehicle treated (top) or 22d treated (bottom) mice. Mice were analyzed 8 weeks after transplantation.
  • FIG. 8 Cartoon depicting proposed model illustrating HDAC8-mediated p53 inactivation contributes to CBF ⁇ -SMMHC-associated AML LSC maintenance.
  • the leukemogenic fusion protein CBF ⁇ -SMMHC recruits p53 and HDAC8 into an aberrant protein complex, thereby inhibiting the tumor suppressor p53 activity is through aberrant deacetylation by HDAC8.
  • Inhibiting HDAC8 deacetylase activity by HDAC8 selective pharmacological inhibitor leads to reactivation of p53 in AML LSCs.
  • This novel p53-inactivating mechanism highlights a promising approach to restore p53 activity, and enhance targeting of AML LSCs.
  • FIG. 9 CM interacts with mostly deacetylated p53 proteins.
  • DUOLINK® in situ PLA in 32D-CM cells using mouse anti-CBF ⁇ , rabbit anti-p53 or Ac-p53 (K379) and PLA probes. Red fluorescent spots indicate CM-p53 or CM-Ac-p53 protein interactions (top), DAPI-stained nucleus is in blue (center) and GFP reporter indicates transduced cells (bottom).
  • FIG. 10 HDAC8 interacts with CM C-terminal region.
  • DUOLINK® in situ PLA in 32D cells expressing FL-CM, deletion mutant ⁇ C95 using mouse anti-CBF ⁇ , xrabbit anti-HDAC8 and PLA probes. Fluorescent spots indicate CM-HDAC8 protein interactions (top), DAPI-stained nucleus is in center, and GFP reporter indicates transduced cells (bottom).
  • FIG. 11 Acetylation of p53 is not affected in CM deletion mutants unable to interact with p53 or HDAC8.
  • FIGS. 12A-12B HDAC8i induces activation of p53 targets in progenitor cells expressing endogenous levels of CM.
  • FIGS. 13A-13B HDAC8i treatment selectively induce of apoptosis and p53 acetylation in inv(16) + AML CD34 + cells.
  • FIG. 14A-14B Confirmation of p53 knock-down by lentivirus expressing shRNA.
  • FIG. 14A Change in p53 expression level in GFP sorted MV4-11 cells transduced with pLKO.1-GFP lentivirus expressing sh-p53 or non-silencing control (sh-ctrl) normalized to levels of b-actin. Shown are mean ⁇ SEM.
  • FIG. 14B Western blot analysis of p53 in GFP sorted MV4-11 cells transduced with pLKO.1-GFP lentivirus expressing sh-p53 or sh-ctrl. Levels of b-actin serve as loading control.
  • FIGS. 15A-15F HDAC8i 22d treatment significantly reduces engraftment and progression of AML.
  • FIG. 15E The frequency of dTomato+ cells in the PB 8 weeks after transplantation of 2 ⁇ 10 6 AML cells treated with
  • FIGS. 16A-16B CBFb-SMMHC does not affect p53 mRNA expression.
  • FIG. 16A Relative expression of p53 mRNA in 32D-Cbfb or 32D-CM cells as determined by qRT-PCR. Shown are mean ⁇ SD.
  • FIG. 16B Relative expression of p53 mRNA in pre-leukemic progenitor subsets sorted from induced Cbfb +/56M Mx1Cre, analyzed by qRT-PCR.
  • Phenotypic progenitor subsets are defined as myeloid progenitors (MPs) (Lin ⁇ /ckit + /Sca1 ⁇ ), common myeloid progenitors (CMPs) (Lin ⁇ /ckit + /Sca1 ⁇ /CD34 + /FcgR lo ), granulocyte-macrophage progenitors (GMPs) (Lin ⁇ /ckit + /Sca1 ⁇ /CD34 + /FcgR hi ), and megakaryocyte-erythroid progenitors (MEPs) (Lin ⁇ /ckit + /Sca1 ⁇ /CD34 + /FcgR hi ).
  • MPs myeloid progenitors
  • CMPs common myeloid progenitors
  • GMPs granulocyte-macrophage progenitors
  • MMPs megakaryocyte-erythroid progenitors
  • FIGS. 17A-17C Effects of 22d on p53 targets are p53-dependent.
  • FIG. 17A Change in p53 expression level in 32D-CM cells transduced with pLKO.1 lentivirus expressing sh-p53 or non-silencing control (sh-Ctrl) normalized to levels of b-actin. Shown are mean ⁇ SEM.
  • FIG. 17B Western blotting of p53 levels in 32D-CM cells expressing sh-p53 or sh-Ctrl. Levels of 3-actin were detected as loading control.
  • FIG. 17C Fold activation of p53 target genes treated with 22d (10 mM) for 16 h, determined by qRT-PCR. Relative expression of each target gene was normalized to levels of ACTB. * P ⁇ 0.05.
  • FIGS. 18A-18F Treatment of HDAC8i 22d does not affect normal HSC engraftment. Engraftment of normal CB CD34 + cells treated with 22d (10 ⁇ M) or vehicle for 48 h into sub-lethally irradiated NOD/SCID/interleukin-2 receptor-g chain deficient (NSG) mice. Shown are engraftment levels of each population (CD45+, CD34+, CD33+, CD14+, CD15+) in the bone marrow and spleen weight at 16 weeks after transplantation. Each dot represents result from an individual mouse and line indicates medium. ns, not significant. FIGS. 18A-18F (in order): CD45+, CD34+, CD33+, CD14+, CD15+, and spleen.
  • FIG. 19 CBFb-SMMHC does not affect HDAC8 expression.
  • FIG. 20 HDAC8i selectively induce acetylated p53 in CM-expressing cells.
  • Levels of ⁇ -actin serve as loading control.
  • FIGS. 21A-21D HDAC8 inhibition activates p53 and induces p53-dependent apoptosis in human AML cells.
  • FIG. 21A Relative survival of human AML cells treated with HDAC8i (Cmpd 22d) for 48 h, as determined by Annexin V labeling and normalized to vehicle-treated controls.
  • FIG. 21B Western blot analysis of Ac-p53, total p53, Ac-H3, Ac-H4, ⁇ -actin in MV4-11 cells treated with indicated doses of HDAC8i for 6 h.
  • FIG. 21C Fold change in mRNA levels of p53 targets after treatment with PCI-48012 (10 mM or 20 mM) for 16 h.
  • 21D Western blot analysis of p53, ⁇ -actin in MV4-11 cells transduced with control or sh-p53 (top). Relative survival of control or sh-p53 transduced MV4-11 cells treated with HDAC8i (bottom).
  • FIGS. 22A-22E Activity of HDAC8i compounds on AML cell proliferation and survival.
  • FIGS. 22A-22C Relative survival of non-p53-mutated AML cell lines (Mv4-11, MOLM13, OCI-AML3; FIGS. 22A-22C , respectively) treated with various HDAC8i (22d, 5b, 5e, 5 h) for 48 h, as determined by Annexin V labeling and normalized to vehicle-treated controls.
  • FIGS. 22D-22E Relative proliferation of AML cell lines (Mv4-11, MOLM13, FIGS. 22D,22E , respectively) treated with various HDAC8i (22d, 5b, 5e, 5h) for 48 h, as determined by Cell Titer-Glo Luminescent Cell Viability Assay and normalized to vehicle treated controls.
  • FIG. 23 Figure depicts histogram of survival rate of PBSC and CB cells showing no significant difference based on cell origin upon contact with Cmpd 22d.
  • FIGS. 24A-24B Histograms demonstrating no significant changes in p53 mRNA or protein levels by CBF ⁇ -SMMHC expression in 32D myeloid progenitor cell line ( FIG. 24A ) or in primary myeloid progenitor cells ( FIG. 24B ). Legend: y-axis: relative expression of indicated proteins. Legend ( FIG. 24B ): Control (filled), pre-leukemic (open).
  • FIG. 25 The figure depicts the effect of agents PCI-24781, PCI-48012 and Nutlin on protein expression for Ac-p53, p53 and ⁇ -actin.
  • FIGS. 26A-26C Effects of HDAC8 inhibitors on interaction between p53 and CM.
  • FIG. 26A Figure depicts effects of Cmpd 22d on IgG and IP expression.
  • FIG. 26B Figure depicts protein expression levels after contact with Cmpd 22d on Ac-p53, p53 and J ⁇ -actin under the indicated timing and wash conditions.
  • FIG. 26C Histogram depicting survival rate with and without wash of Cmpd 22d.
  • FIGS. 27A-27B Inhibition of HDAC8 selectively activate p53 in inv(16)+ AML CD34+ cells.
  • FIG. 27A Figure depicts Western blotting of Ac-p53, (K382), and p53 levels in inv(16)+ AML CD34+ cells upon contact with Cmpd 22d.
  • FIG. 27B Histogram depicts the fold activation of the indicated p53 target genes (in order left to right: p21, hdm2, 14-3-3 ⁇ , puma) in inv(16)+ AML CD34+ and normal CD34+ cells. Legend: inv(16)+ AML CD34+ (black filled); normal CD34+ cells (gray filled).
  • substituent groups are specified by their conventional chemical formulae, written from left to right, they equally encompass the chemically identical substituents that would result from writing the structure from right to left, e.g., —CH 2 O— is equivalent to —OCH 2 —.
  • alkyl by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e., unbranched) or branched chain, or combination thereof, which may be fully saturated, mono- or polyunsaturated and can include di- and multivalent radicals, having the number of carbon atoms designated (i.e., C 1 -C 10 means one to ten carbons). Alkyl is not cyclized.
  • saturated hydrocarbon radicals include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, (cyclohexyl)methyl, homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.
  • An unsaturated alkyl group is one having one or more double bonds or triple bonds.
  • unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs and isomers.
  • An alkoxy is an alkyl attached to the remainder of the molecule via an oxygen linker (—O—).
  • alkylene by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkyl, as exemplified, but not limited by, —CH 2 CH 2 CH 2 CH 2 —. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms.
  • a “lower alkyl” or “lower alkylene” is a shorter chain alkyl or alkylene group, generally having eight or fewer carbon atoms.
  • heteroalkyl by itself or in combination with another term, means, unless otherwise stated, a stable straight or branched chain, or combinations thereof, consisting of at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, S, Se and Si, and wherein the nitrogen, selenium, and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. Heteroalkyl is not cyclized. The heteroatom(s) O, N, P, S, Se, and Si may be placed at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule.
  • Examples include, but are not limited to: —CH 2 —CH 2 —O—CH 3 , —CH 2 —CH 2 —NH—CH 3 , —CH 2 —CH 2 —N(CH 3 )—CH 3 , —CH 2 —S—CH 2 —CH 3 , —CH 2 —CH 2 , —S(O)—CH 3 , —CH 2 —CH 2 —S(O) 2 —CH 3 , —CH ⁇ CH—O—CH 3 , —Si(CH 3 ) 3 , —CH 2 —CH ⁇ N—OCH 3 , —CH ⁇ CH—N(CH 3 )—CH 3 , —O—CH 3 , —O—CH 2 —CH 3 , and —CN.
  • Up to two heteroatoms may be consecutive, such as, for example, —CH 2 —NH—OCH 3 .
  • heteroalkylene by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from heteroalkyl, as exemplified, but not limited by, —CH 2 —CH 2 —S—CH 2 —CH 2 — and —CH 2 —S—CH 2 —CH 2 —NH—CH 2 —.
  • heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like).
  • heteroalkyl groups include those groups that are attached to the remainder of the molecule through a heteroatom, such as —C(O)R′, —C(O)NR′, —NR′R′′, —OR′, —SeR′, —SR′, and/or —SO 2 R′.
  • heteroalkyl is recited, followed by recitations of specific heteroalkyl groups, such as —NR′R′′ or the like, it will be understood that the terms heteroalkyl and —NR′R′′ are not redundant or mutually exclusive. Rather, the specific heteroalkyl groups are recited to add clarity. Thus, the term “heteroalkyl” should not be interpreted herein as excluding specific heteroalkyl groups, such as —NR′R′′ or the like.
  • cycloalkyl and heterocycloalkyl mean, unless otherwise stated, cyclic versions of“alkyl” and “heteroalkyl,” respectively. Cycloalkyl and heterocycloalkyl are not aromatic. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like.
  • heterocycloalkyl examples include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like.
  • a “cycloalkylene” and a “heterocycloalkylene,” alone or as part of another substituent, means a divalent radical derived from a cycloalkyl and heterocycloalkyl, respectively.
  • aryl means, unless otherwise stated, a polyunsaturated, aromatic, hydrocarbon substituent, which can be a single ring or multiple rings (e.g. 1 to 3 rings) that are fused together (i.e., a fused ring aryl) or linked covalently.
  • a fused ring aryl refers to multiple rings fused together wherein at least one of the fused rings is an aryl ring.
  • heteroaryl refers to aryl groups (or rings) that contain at least one heteroatom (e.g. N, O, or S), wherein sulfur heteroatoms are optionally oxidized, and the nitrogen heteroatoms are optionally quaternized.
  • heteroaryl includes fused ring heteroaryl groups (i.e., multiple rings fused together wherein at least one of the fused rings is a heteroaromatic ring).
  • a 5,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 5 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring.
  • a 6,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring.
  • a 6,5-fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members and the other ring has 5 members, and wherein at least one ring is a heteroaryl ring.
  • a heteroaryl group can be attached to the remainder of the molecule through a carbon or heteroatom.
  • Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinoly
  • arylene and heteroarylene are selected from the group of acceptable substituents described below.
  • a fused ring heterocycloalkyl-aryl is an aryl fused to a heterocycloalkyl.
  • a fused ring heterocycloalkyl-heteroaryl is a heteroaryl fused to a heterocycloalkyl.
  • a fused ring heterocycloalkyl-cycloalkyl is a heterocycloalkyl fused to a cycloalkyl.
  • a fused ring heterocycloalkyl-heterocycloalkyl is a heterocycloalkyl fused to another heterocycloalkyl.
  • Fused ring heterocycloalkyl-aryl, fused ring heterocycloalkyl-heteroaryl, fused ring heterocycloalkyl-cycloalkyl, or fused ring heterocycloalkyl-heterocycloalkyl may each independently be unsubstituted or substituted with one or more of the substituents described herein.
  • Spirocyclic rings are two or more rings wherein adjacent rings are attached through a single atom.
  • the individual rings within spirocyclic rings may be identical or different. Individual rings in spirocyclic rings may be substituted or unsubstituted and may have different substituents from other individual rings within a set of spirocyclic rings.
  • Spirocylic rings may be substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heterocycloalkylene and individual rings within a spirocyclic ring group may be any of the immediately previous list, including having all rings of one type (e.g.
  • heterocyclic spirocyclic rings means a spirocyclic rings wherein at least one ring is a heterocyclic ring and wherein each ring may be a different ring.
  • substituted spirocyclic rings means that at least one ring is substituted and each substituent may optionally be different.
  • halo or “halogen,” by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as “haloalkyl” are meant to include monohaloalkyl and polyhaloalkyl.
  • halo(C 1 -C 4 )alkyl includes, but is not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.
  • acyl means, unless otherwise stated, —C(O)R where R is a substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • oxo means an oxygen that is double bonded to a carbon atom.
  • Substituents for the alkyl and heteroalkyl radicals can be one or more of a variety of groups selected from, but not limited to, —OR′, ⁇ O, ⁇ NR′, ⁇ N—OR′, —NR′R′′, —SR′, -halogen, —SiR′R′′R′′′, —OC(O)R′, —C(O)R′, —CO 2 R′, —CONR′R′′, —OC(O)NR′R′′, —NR′′C(O)R′, —NR′—C(O)NR′′R′′′, —NR′′C(O) 2 R′, —NR—C(NR′R′′R′′′) ⁇ NR′′′′, —NR—C(NR′R′′R′′′) ⁇ NR′′′′,
  • R′, R′′, R′′′, and R′′′′ each preferably independently refer to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g., aryl substituted with 1-3 halogens), substituted or unsubstituted alkyl, alkoxy, or thioalkoxy groups, or arylalkyl groups.
  • each of the R groups is independently selected as are each R′, R′′, R′′′, and R′′′′ group when more than one of these groups is present.
  • R′ and R′′ When R′ and R′′ are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring.
  • —NR′R′′ includes, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl.
  • alkyl is meant to include groups including carbon atoms bound to groups other than hydrogen groups, such as haloalkyl (e.g., —CF 3 and —CH 2 CF 3 ) and acyl (e.g., —C(O)CH 3 , —C(O)CF 3 , —C(O)CH 2 OCH 3 , and the like).
  • substituents for the aryl and heteroaryl groups are varied and are selected from, for example: —OR′, —NR′R′′, —SR′, -halogen, —SiR′R′′R′′′, —OC(O)R′, —C(O)R′, —CO 2 R′, —CONR′R′′, —OC(O)NR′R′′, —NR′′C(O)R′, —NR′—C(O)NR′′R′′′, —NR′′C(O) 2 R′, —NR—C(NR′R′′R′′′) ⁇ NR′′′′, —NR—C(NR′R′′) ⁇ NR′′′, —S(O)R′, —S(O) 2 R′, —S(O) 2 NR′R′′, —NRSO 2 R′, —CN, —NO 2 , —R′, —N 3 , —CH(Ph
  • Substituents for rings may be depicted as substituents on the ring rather than on a specific atom of a ring (commonly referred to as a floating substituent).
  • the substituent may be attached to any of the ring atoms (obeying the rules of chemical valency) and in the case of fused rings or spirocyclic rings, a substituent depicted as associated with one member of the fused rings or spirocyclic rings (a floating substituent on a single ring), may be a substituent on any of the fused rings or spirocyclic rings (a floating substituent on multiple rings).
  • the multiple substituents may be on the same atom, same ring, different atoms, different fused rings, different spirocyclic rings, and each substituent may optionally be different.
  • a point of attachment of a ring to the remainder of a molecule is not limited to a single atom (a floating substituent)
  • the attachment point may be any atom of the ring and in the case of a fused ring or spirocyclic ring, any atom of any of the fused rings or spirocyclic rings while obeying the rules of chemical valency.
  • a ring, fused rings, or spirocyclic rings contain one or more ring heteroatoms and the ring, fused rings, or spirocyclic rings are shown with one more floating substituents (including, but not limited to, points of attachment to the remainder of the molecule), the floating substituents may be bonded to the heteroatoms.
  • the ring heteroatoms are shown bound to one or more hydrogens (e.g. a ring nitrogen with two bonds to ring atoms and a third bond to a hydrogen) in the structure or formula with the floating substituent, when the heteroatom is bonded to the floating substituent, the substituent will be understood to replace the hydrogen, while obeying the rules of chemical valency.
  • Two or more substituents may optionally be joined to form aryl, heteroaryl, cycloalkyl, or heterocycloalkyl groups.
  • Such so-called ring-forming substituents are typically, though not necessarily, found attached to a cyclic base structure.
  • the ring-forming substituents may be attached to adjacent members of the base structure.
  • two ring-forming substituents attached to adjacent members of a cyclic base structure create a fused ring structure.
  • the ring-forming substituents may be attached to a single member of the base structure.
  • two ring-forming substituents attached to a single member of a cyclic base structure create a spirocyclic structure.
  • the ring-forming substituents may be attached to non-adjacent members of the base structure.
  • Two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally form a ring of the formula -T-C(O)—(CRR′) q —U—, wherein T and U are independently —NR—, —O—, —CRR′—, or a single bond, and q is an integer of from 0 to 3.
  • two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -A-(CH 2 ) r —B—, wherein A and B are independently —CRR′—, —O—, —NR—, —S—, —S(O)—, —S(O) 2 —, —S(O) 2 NR′—, or a single bond, and r is an integer of from 1 to 4.
  • One of the single bonds of the new ring so formed may optionally be replaced with a double bond.
  • two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula —(CRR′) s —X′—(C′′R′′′) d —, where s and d are independently integers of from 0 to 3, and X′ is —O—, —NR′—, —S—, —S(O)—, —S(O) 2 —, or —S(O) 2 NR′—.
  • R, R′, R′′, and R′′′ are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.
  • heteroatom or “ring heteroatom” are meant to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).
  • a “substituent group,” as used herein, means a group selected from the following moieties:
  • a “size-limited substituent” or “size-limited substituent group,” as used herein, means a group selected from all of the substituents described above for a “substituent group,” wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C 1 -C 20 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C 3 -C 8 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 8 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C 3 -C 8 aryl, and each substituted or unsubstituted heteroaryl is
  • a “lower substituent” or “lower substituent group,” as used herein, means a group selected from all of the substituents described above for a “substituent group,” wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C 1 -C 8 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C 3 -C 7 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C 3 -C 7 aryl, and each substituted or unsubstituted heteroaryl is a substitute
  • Each substituted group described in the compounds herein may be substituted with at least one substituent group. More specifically, each substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and/or substituted heteroarylene described in the compounds herein may be substituted with at least one substituent group. At least one or all of these groups may be substituted with at least one size-limited substituent group. At least one or all of these groups may be substituted with at least one lower substituent group.
  • Each substituted or unsubstituted alkyl may be a substituted or unsubstituted C 1 -C 20 alkyl
  • each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20 membered heteroalkyl
  • each substituted or unsubstituted cycloalkyl may be a substituted or unsubstituted C 3 -C 8 cycloalkyl
  • each substituted or unsubstituted heterocycloalkyl may be a substituted or unsubstituted 3 to 8 membered heterocycloalkyl.
  • Each substituted or unsubstituted alkylene may be a substituted or unsubstituted C 1 -C 20 alkylene
  • each substituted or unsubstituted heteroalkylene may be a substituted or unsubstituted 2 to 20 membered heteroalkylene
  • each substituted or unsubstituted cycloalkylene may be a substituted or unsubstituted C 3 -C 8 cycloalkylene
  • each substituted or unsubstituted heterocycloalkylene may be a substituted or unsubstituted 3 to 8 membered heterocycloalkylene
  • each substituted or unsubstituted arylene may be a substituted or unsubstituted C 3 -C 8 arylene
  • each substituted or unsubstituted heteroaryl may be a substituted or unsubstituted C 3 -C 8 heteroarylene.
  • Each substituted or unsubstituted alkyl may be a substituted or unsubstituted C 1 -C 8 alkyl
  • each substituted or unsubstituted heteroalkyl may be a substituted or unsubstituted 2 to 8 membered heteroalkyl
  • each substituted or unsubstituted cycloalkyl may be a substituted or unsubstituted C 3 -C 7 cycloalkyl
  • each substituted or unsubstituted heterocycloalkyl may be a substituted or unsubstituted 3 to 7 membered heterocycloalkyl
  • each substituted or unsubstituted aryl may be a substituted or unsubstituted C 3 -C 7 aryl
  • each substituted or unsubstituted heteroaryl may be a substituted or unsubstituted C 3 -C 7 heteroaryl.
  • Each substituted or unsubstituted alkylene may be a substituted or unsubstituted C 1 -C 8 alkylene
  • each substituted or unsubstituted heteroalkylene may be a substituted or unsubstituted 2 to 8 membered heteroalkylene
  • each substituted or unsubstituted cycloalkylene may be a substituted or unsubstituted C 3 -C 7 cycloalkylene
  • each substituted or unsubstituted heterocycloalkylene may be a substituted or unsubstituted 3 to 7 membered heterocycloalkylene
  • each substituted or unsubstituted arylene may be a substituted or unsubstituted C 3 -C 7 arylene
  • each substituted or unsubstituted heteroarylene may be a substituted or unsubstituted C 3 -C 7 heteroarylene.
  • Certain compounds described herein possess asymmetric carbon atoms (optical or chiral centers) or double bonds; the enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids, and individual isomers are encompassed within the scope of the compounds described herein.
  • the compounds described herein do not include those that are known in art to be too unstable to synthesize and/or isolate.
  • Compounds described herein include compounds in racemic and optically pure forms.
  • Optically active (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.
  • the compounds described herein contain olefinic bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers.
  • isomers refers to compounds having the same number and kind of atoms, and hence the same molecular weight, but differing in respect to the structural arrangement or configuration of the atoms.
  • tautomer refers to one of two or more structural isomers which exist in equilibrium and which are readily converted from one isomeric form to another.
  • structures depicted herein are also meant to include all stereochemical forms of the structure; i.e., the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope described herein.
  • structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms.
  • compounds described herein include the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by 13 C- or 14 C-enriched carbon.
  • the compounds described herein may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds.
  • the compounds may be radiolabeled with radioactive isotopes, such as for example tritium ( 3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C).
  • radioactive isotopes such as for example tritium ( 3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C).
  • Compounds described herein further include all isotopic variations thereof, whether radioactive or not.
  • an analog or “analogue” is used in accordance with its plain ordinary meaning within Chemistry and Biology and refers to a chemical compound that is structurally similar to another compound (i.e., a so-called “reference” compound) but differs in composition, e.g., in the replacement of one atom by an atom of a different element, or in the presence of a particular functional group, or the replacement of one functional group by another functional group, or the absolute stereochemistry of one or more chiral centers of the reference compound. Accordingly, an analog is a compound that is similar or comparable in function and appearance but not in structure or origin to a reference compound.
  • a or “an,” as used in herein means one or more.
  • substituted with a[n] means the specified group may be substituted with one or more of any or all of the named substituents.
  • a group such as an alkyl or heteroaryl group, is “substituted with an unsubstituted C 1 -C 20 alkyl, or unsubstituted 2 to 20 membered heteroalkyl,” the group may contain one or more unsubstituted C 1 -C 20 alkyls, and/or one or more unsubstituted 2 to 20 membered heteroalkyls.
  • R-substituted where a moiety is substituted with an R substituent, the group may be referred to as “R-substituted.” Where a moiety is R-substituted, the moiety is substituted with at least one R substituent and each R substituent is optionally different. Where a particular R group is present in the description of a chemical genus (such as Formula (I)), a decimal symbol may be used to distinguish each appearance of that particular R group. For example, where multiple R 13 substituents are present, each R 13 substituent may be distinguished as R 13.1 , R 13.2 , R 13.3 , R 13.4 , etc., wherein each of R 13.1 , R 13.2 , R 13.3 , R 13.4 , etc. is defined within the scope of the definition of R 13 and optionally differently.
  • DNA and RNA refer to deoxyribonucleic acid and ribonucleic acid, respectively.
  • Nucleic acid refers to deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or double-stranded form, and complements thereof.
  • polynucleotide refers to a linear sequence of nucleotides.
  • nucleotide typically refers to a single unit of a polynucleotide, i.e., a monomer. Nucleotides can be ribonucleotides, deoxyribonucleotides, or modified versions thereof.
  • nucleic acid as used herein also refers nucleic acids that have the same basic chemical structure as naturally occurring nucleic acids. Such analogues have modified sugars and/or modified ring substituents, but retain the same basic chemical structure as the naturally occurring nucleic acid.
  • a nucleic acid mimetic refers to chemical compounds that have a structure that is different the general chemical structure of a nucleic acid, but functions in a manner similar to a naturally occurring nucleic acid.
  • Examples of such analogues include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, and peptide-nucleic acids (PNAs).
  • Synthetic mRNA refers to any mRNA derived through non-natural means such as standard oligonucleotide synthesis techniques or cloning techniques. Such mRNA may also include non-proteinogenic derivatives of naturally occurring nucleotides. Additionally, “synthetic mRNA” herein also includes mRNA that has been expressed through recombinant techniques or exogenously, using any expression vehicle, including but not limited to prokaryotic cells, eukaryotic cell lines, and viral methods. “Synthetic mRNA” includes such mRNA that has been purified or otherwise obtained from an expression vehicle or system.
  • polypeptide “peptide” and “protein” are used interchangeably herein to refer to a polymer of amino acid residues, wherein the polymer may optionally be conjugated to a moiety that does not consist of amino acids.
  • the terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymer.
  • peptidyl and “peptidyl moiety” means a monovalent peptide.
  • amino acid refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids.
  • Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, ⁇ -carboxyglutamate, and O-phosphoserine.
  • Amino acid analogs refers to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid.
  • Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid.
  • Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single-letter codes.
  • “Conservatively modified variants” applies to both amino acid and nucleic acid sequences. With respect to particular nucleic acid sequences, conservatively modified variants refers to those nucleic acids which encode identical or essentially identical amino acid sequences, or where the nucleic acid does not encode an amino acid sequence, to essentially identical sequences. Because of the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For instance, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at every position where an alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide.
  • nucleic acid variations are “silent variations,” which are one species of conservatively modified variations. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic acid.
  • each codon in a nucleic acid except AUG, which is ordinarily the only codon for methionine, and TGG, which is ordinarily the only codon for tryptophan
  • TGG which is ordinarily the only codon for tryptophan
  • amino acid sequences one of skill will recognize that individual substitutions, deletions or additions to a nucleic acid, peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a “conservatively modified variant” where the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to and do not exclude polymorphic variants, interspecies homologs, and alleles described herein.
  • the following eight groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), Glycine (G); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); 7) Serine (S), Threonine (T); and 8) Cysteine (C), Methionine (M) (see. e.g., Creighton, Proteins (1984)).
  • pharmaceutically acceptable salts is meant to include salts of the active compounds that are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein.
  • base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent.
  • pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt.
  • acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent.
  • Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfiuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, oxalic, methanesulfonic, and the like.
  • inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfiuric, hydr
  • salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19).
  • Certain specific compounds described herein contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.
  • the compounds described herein may exist as salts, such as with pharmaceutically acceptable acids.
  • the compounds described herein include such salts.
  • Non-limiting examples of such salts include hydrochlorides, hydrobromides, phosphates, sulfates, methanesulfonates, nitrates, maleates, acetates, citrates, fumarates, propionates, tartrates (e.g., (+)-tartrates, ( ⁇ )-tartrates, or mixtures thereof including racemic mixtures), succinates, benzoates, and salts with amino acids such as glutamic acid, and quaternary ammonium salts (e.g. methyl iodide, ethyl iodide, and the like). These salts may be prepared by methods known to those skilled in the art.
  • the neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner.
  • the parent form of the compound may differ from the various salt forms in certain physical properties, such as solubility in polar solvents.
  • the compounds described herein may be provided in a prodrug form.
  • Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide their respective active forms.
  • Prodrugs of the compounds described herein may be converted in vivo after administration.
  • prodrugs can be converted to the compounds described herein by chemical or biochemical methods in an ex vivo environment, such as, for example, when contacted with a suitable enzyme or chemical reagent.
  • Certain compounds described herein can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms compounds described herein are equivalent to unsolvated forms. Certain compounds described herein may exist in multiple crystalline or amorphous forms. In general, all physical forms compounds described herein are equivalent for their uses described herein.
  • “Pharmaceutically acceptable excipient” and “pharmaceutically acceptable carrier” refer to a substance that aids the administration of an active agent to and absorption by a subject and can be included in the compositions described herein without causing a significant adverse toxicological effect on the patient.
  • Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, lactated Ringer's, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinyl pyrrolidine, and colors, and the like.
  • Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with the compounds described herein.
  • auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with the compounds described herein.
  • auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with the compounds described herein.
  • auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents
  • preparation is intended to include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component with or without other carriers, is surrounded by a carrier, which is thus in association with it.
  • carrier providing a capsule in which the active component with or without other carriers, is surrounded by a carrier, which is thus in association with it.
  • cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.
  • administering means oral administration, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal or subcutaneous administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to a subject.
  • Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal).
  • Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial.
  • Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc.
  • compositions disclosed herein can be delivered by transdermally, by a topical route, formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols.
  • Oral preparations include tablets, pills, powder, dragees, capsules, liquids, lozenges, cachets, gels, syrups, slurries, suspensions, etc., suitable for ingestion by the patient.
  • Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules.
  • Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water/propylene glycol solutions.
  • the compositions of the present invention may additionally include components to provide sustained release and/or comfort.
  • Such components include high molecular weight, anionic mucomimetic polymers, gelling polysaccharides and finely-divided drug carrier substrates. These components are discussed in greater detail in U.S. Pat. Nos. 4,911,920; 5,403,841; 5,212,162; and 4,861,760. The entire contents of these patents are incorporated herein by reference in their entirety for all purposes.
  • the compositions disclosed herein can also be delivered as microspheres for slow release in the body.
  • microspheres can be administered via intradermal injection of drug-containing microspheres, which slowly release subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645, 1995; as biodegradable and injectable gel formulations (see, e.g., Gao Pharm. Res. 12:857-863, 1995); or, as microspheres for oral administration (see, e.g., Eyles, J. Pharm. Pharmacol. 49:669-674, 1997).
  • the formulations of the compositions of the present invention can be delivered by the use of liposomes which fuse with the cellular membrane or are endocytosed, i.e., by employing receptor ligands attached to the liposome, that bind to surface membrane protein receptors of the cell resulting in endocytosis.
  • liposomes particularly where the liposome surface carries receptor ligands specific for target cells, or are otherwise preferentially directed to a specific organ, one can focus the delivery of the compositions of the present invention into the target cells in vivo.
  • the compositions can also be delivered as nanoparticles.
  • compositions may include compositions wherein the active ingredient (e.g. compounds described herein, including embodiments or examples) is contained in a therapeutically effective amount, i.e., in an amount effective to achieve its intended purpose.
  • a therapeutically effective amount i.e., in an amount effective to achieve its intended purpose.
  • the actual amount effective for a particular application will depend, inter alia. on the condition being treated.
  • compositions When administered in methods to treat a disease, such compositions will contain an amount of active ingredient effective to achieve the desired result, e.g., modulating the activity of a target molecule, and/or reducing, eliminating, or slowing the progression of disease symptoms.
  • the dosage and frequency (single or multiple doses) administered to a mammal can vary depending upon a variety of factors, for example, whether the mammal suffers from another disease, and its route of administration; size, age, sex, health, body weight, body mass index, and diet of the recipient; nature and extent of symptoms of the disease being treated, kind of concurrent treatment, complications from the disease being treated or other health-related problems.
  • Other therapeutic regimens or agents can be used in conjunction with the methods and compounds of Applicants' invention. Adjustment and manipulation of established dosages (e.g., frequency and duration) are well within the ability of those skilled in the art.
  • the compounds and complexes described herein can be used in combination with one another, with other active drugs known to be useful in treating a disease (e.g. anti-cancer drugs) or with adjunctive agents that may not be effective alone, but may contribute to the efficacy of the active agent.
  • active drugs e.g. anti-cancer drugs
  • adjunctive agents may not be effective alone, but may contribute to the efficacy of the active agent.
  • co-administer it is meant that a composition described herein is administered at the same time, just prior to, or just after the administration of one or more additional therapies, for example an anticancer agent as described herein.
  • the compound of the invention can be administered alone or can be co-administered to the patient.
  • Co-administration is meant to include simultaneous or sequential administration of the compound individually or in combination (more than one compound or agent).
  • the preparations can also be combined, when desired, with other active substances (e.g. anticancer agents).
  • Co-administration includes administering one active agent (e.g. a complex described herein) within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of a second active agent (e.g. anti-cancer agents). Also contemplated herein, are embodiments, where co-administration includes administering one active agent within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of a second active agent. Co-administration includes administering two active agents simultaneously, approximately simultaneously (e.g., within about 1, 5, 10, 15, 20, or 30 minutes of each other), or sequentially in any order. In embodiments, co-administration can be accomplished by co-formulation, i.e., preparing a single pharmaceutical composition including both active agents. In other embodiments, the active agents can be formulated separately. In embodiments, the active and/or adjunctive agents may be linked or conjugated to one another. In embodiments, the compounds and complexes described herein may be combined with treatments for cancer such as chemotherapy or radiation therapy.
  • a second active agent e.g. anti-cancer agents.
  • HDAC8 inhibitor and “HDAC8i” are used interchangeably herein and refer a composition (e.g. compound, peptide, protein, nucleic acid, or antibody) which reduces the activity of HDAC8 (Histone Deacetylase 8) relative to the activity of HDAC8 in the absence of the inhibitor.
  • HDAC8 inhibitors may be selective for HDAC8 as described herein.
  • the HDAC8 inhibitor may be a HDAC8 inhibitor compound (e.g. a compound having a molecular weight (MW) of less than about 1000 Da).
  • the HDAC8 inhibitor compound may be a compound described herein.
  • HDAC8 inhibitor compounds further include compounds known to selectively inhibit HDAC8 expression or activity including one or more of those exemplified in, for example, U.S. Pat. No. 7,820,711; PCT/JP2011/050647; and/or PCT/US2014/012968.
  • HDAC8 inhibitor compounds include one or more of the compounds described herein, and further includes, for example, one or more of the compounds described by K. Krennhrubec, et al., Bioorg. Med. Chem. Lett. 2007, 17, 2874-2878; P. Galletti, et al., ChemMedChem. 2009, 4, 1991-2001; E. Hu, et al., J. Pharmacol. Exp. Ther. 2003, 307, 720-728; W. Tang, et al., Bioorg. Med. Chem. Lett. 2011, 21, 2601-2605; S. Balasubramanian, et al., Leukemia 2008, 22, 1026-1034; L. Whitehead, et al., Bioorg. Med. Chem. 2011, 19, 4626-4634; and T. Suzuki, et al., ChemMedChem. 2014, 9, 657-664.
  • the HDAC8 inhibitor may be a HDAC8 inhibitor antibody (e.g. those described by PCT/US2000/033622).
  • the HDAC8 inhibitor may be a HDAC8 inhibitor polynucleotide.
  • the HDAC8 inhibitor polynucleotide may be a mdRNA as described by, for example, PCT/US2008/055612.
  • the HDAC8 inhibitor polynucleotide may be RNA (e.g. a HDAC8 inhibitor RNA), siRNA (e.g. a HDAC8 inhibitor siRNA), shRNA (e.g. a HDAC8 inhibitor shRNA) or a miRNA (e.g. a HDAC8 inhibitor miRNA).
  • the HDAC8 inhibitor may be a HDAC8 inhibitor protein.
  • a selective HDAC8 inhibitor described herein may have an IC 50 for HDAC8 activity that is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 fold lower than the IC 50 for one or more of HDAC1, HDAC2, HDAC3, HDAC6, HDAC10, and/or HDAC 11.
  • a selective HDAC8 inhibitor may have an IC 50 for HDAC8 acetyltransferase activity that is about 5, 10, 50, 150, 200, 250, 300, 350, 400, 450 or more than about 500 fold lower than the IC 50 for acetyltransferase activity of another HDAC (e.g. HDAC1, HDAC2, HDAC3, HDAC6, HDAC10, or HDAC 11).
  • Specific”, “specifically”, “specificity”, or the like of a compound refers to the compound's ability to cause a particular action, such as inhibition, to a particular molecular target with minimal or no action to other proteins in the cell.
  • HDAC8 is used herein and according to its common, ordinary meaning and refers to proteins of the same or similar names and functional fragments and homologs thereof.
  • the term includes any recombinant or naturally occurring form of HDAC8 (e.g. Histone deacetylase 8; GI No: 8132878), or variants or fragments thereof that maintain HDAC8 activity (e.g. within at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% activity compared to HDAC8).
  • p53 is used herein and according to its common, ordinary meaning and refers to proteins of the same or similar names and functional fragments and homologs thereof.
  • the term includes any recombinant or naturally occurring form of p53 (e.g. GI No: 23491729), or variants or fragments thereof that maintain p53 activity (e.g. within at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% activity compared to p53).
  • a “mutated p53” is a p53 variant that is aberrantly acetylated or deacetylated resulting from a mutation to the wildtype p53 amino acid sequence.
  • non-mutated p53 refers to p53 variants which are correctly acetylated or deacetylated.
  • a non-mutated p53 may include mutations so long as those mutations impart no effect on p53 acetylation or deacetylation.
  • a “non-mutated p53 cancer” refers to a cancer characterized by correctly acetylated or deacetylated p53.
  • a “mutated p53 cancer” refers to a cancer characterized by incorrectly acetylated or deacetylated p53.
  • treating refers to any indicia of success in the treatment or amelioration of an injury, disease, pathology or condition, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the injury, pathology or condition more tolerable to the patient; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; improving a patient's physical or mental well-being.
  • the treatment or amelioration of symptoms can be based on objective or subjective parameters; including the results of a physical examination, neuropsychiatric exams, and/or a psychiatric evaluation.
  • the term “treating” and conjugations thereof, include prevention of an injury, pathology, condition, or disease.
  • an “effective amount” is an amount sufficient to accomplish a stated purpose (e.g. achieve the effect for which it is administered, treat a disease, reduce enzyme activity, increase enzyme activity, reduce one or more symptoms of a disease or condition).
  • An example of an “effective amount” is an amount sufficient to contribute to the treatment, prevention, or reduction of a symptom or symptoms of a disease, which could also be referred to as a “therapeutically effective amount.”
  • a “reduction” of a symptom or symptoms means decreasing of the severity or frequency of the symptom(s), or elimination of the symptom(s).
  • a “prophylactically effective amount” of a drug is an amount of a drug that, when administered to a subject, will have the intended prophylactic effect, e.g., preventing or delaying the onset (or reoccurrence) of an injury, disease, pathology or condition, or reducing the likelihood of the onset (or reoccurrence) of an injury, disease, pathology, or condition, or their symptoms.
  • the full prophylactic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses.
  • a prophylactically effective amount may be administered in one or more administrations.
  • the therapeutically effective amount can be initially determined from cell culture assays.
  • Target concentrations will be those concentrations of active compound(s) that are capable of achieving the methods described herein, as measured using the methods described herein or known in the art.
  • therapeutically effective amounts for use in humans can also be determined from animal models.
  • a dose for humans can be formulated to achieve a concentration that has been found to be effective in animals.
  • the dosage in humans can be adjusted by monitoring compounds effectiveness and adjusting the dosage upwards or downwards, as described above. Adjusting the dose to achieve maximal efficacy in humans based on the methods described above and other methods is well within the capabilities of the ordinarily skilled artisan.
  • Dosages may be varied depending upon the requirements of the patient and the compound being employed.
  • the dose administered to a patient, in the context of the present invention should be sufficient to effect a beneficial therapeutic response in the patient over time.
  • the size of the dose also will be determined by the existence, nature, and extent of any adverse side-effects. Determination of the proper dosage for a particular situation is within the skill of the practitioner. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under circumstances is reached.
  • Dosage amounts and intervals can be adjusted individually to provide levels of the administered compound effective for the particular clinical indication being treated. This will provide a therapeutic regimen that is commensurate with the severity of the individual's disease state.
  • an effective prophylactic or therapeutic treatment regimen can be planned that does not cause substantial toxicity and yet is effective to treat the clinical symptoms demonstrated by the particular patient.
  • This planning should involve the careful choice of active compound by considering factors such as compound potency, relative bioavailability, patient body weight, presence and severity of adverse side effects, preferred mode of administration and the toxicity profile of the selected agent.
  • Control or “control experiment” is used in accordance with its plain ordinary meaning and refers to an experiment in which the subjects or reagents of the experiment are treated as in a parallel experiment except for omission of a procedure, reagent, or variable of the experiment. In some instances, the control is used as a standard of comparison in evaluating experimental effects.
  • a control may be the measurement of the activity of a protein in the absence of a compound as described herein.
  • test compound refers to an experimental compound used in a screening process to identify activity, non-activity, or other modulation of a particularized biological target or pathway.
  • modulation refers to a composition that increases or decreases the level of a target molecule or the function of a target molecule or the physical state of the target of the molecule.
  • Modulation refers to the process of changing or varying one or more properties. For example, as applied to the effects of a modulator on a biological target, to modulate means to change by increasing or decreasing a property or function of the biological target or the amount of the biological target.
  • inhibition means negatively affecting (e.g. decreasing) the activity or function of the protein relative to the activity or function of the protein in the absence of the inhibitor.
  • Inhibition may refer to reduction of a disease or symptoms of disease.
  • Inhibition may refer to a reduction in the activity of a particular protein or nucleic acid target.
  • inhibition includes, at least in part, partially or totally blocking stimulation, decreasing, preventing, or delaying activation, or inactivating, desensitizing, or down-regulating signal transduction or enzymatic activity or the amount of a protein.
  • Contacting is used in accordance with its plain ordinary meaning and refers to the process of allowing at least two distinct species (e.g. chemical compounds including biomolecules or cells) to become sufficiently proximal to react, interact or physically touch. It should be appreciated; however, the resulting reaction product can be produced directly from a reaction between the added reagents or from an intermediate from one or more of the added reagents that can be produced in the reaction mixture.
  • species e.g. chemical compounds including biomolecules or cells
  • contacting may include allowing two species to react, interact, or physically touch, wherein the two species may be a compound as described herein and a protein or enzyme.
  • Contacting may include allowing a compound described herein to interact with a protein or enzyme that is involved in a signaling pathway.
  • “Patient,” “subject,” “patient in need thereof,” and “subject in need thereof” are herein used interchangeably and refer to a living organism suffering from or prone to a disease or condition that can be treated by administration of a pharmaceutical composition as provided herein.
  • Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, monkeys, goat, sheep, cows, deer, and other non-mammalian animals.
  • a patient may be human.
  • Disease or “condition” or “disorder” refers to a state of being or health status of a patient or subject capable of being treated with the compounds, drugs, pharmaceutical compositions, or methods provided herein.
  • the disease may be a disease related to (e.g. caused by) an abnormal cell growth or abnormal protein activity (e.g. cancer).
  • cancer refers to all types of cancer, neoplasm, or malignant or benign tumors found in mammals, including leukemia, carcinomas and sarcomas.
  • exemplary cancers include acute myeloid leukemia (“AML”), chronic myelogenous leukemia (“CML”), and cancer of the brain, breast, pancreas, colon, liver, kidney, lung, non-small cell lung, melanoma, ovary, sarcoma, and prostate.
  • AML acute myeloid leukemia
  • CML chronic myelogenous leukemia
  • cancer of the brain breast, pancreas, colon, liver, kidney, lung, non-small cell lung, melanoma, ovary, sarcoma, and prostate.
  • Additional examples include, cervix cancers, stomach cancers, head & neck cancers, uterus cancers, mesothelioma, metastatic bone cancer, Medulloblastoma, Hodgkin's Disease, Non-Hodgkin's Lymphoma, multiple myeloma, neuroblastoma, ovarian cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, primary brain tumors, cancer, malignant pancreatic insulanoma, malignant carcinoid, urinary bladder cancer, premalignant skin lesions, testicular cancer, lymphomas, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, endometrial cancer, adrenal cortical cancer, and neoplasms of the endocrine and exocrine pancreas.
  • leukemia refers broadly to progressive, malignant diseases of the blood-forming organs and is generally characterized by a distorted proliferation and development of leukocytes and their precursors in the blood and bone marrow. Leukemia is generally clinically classified on the basis of (1) the duration and character of the disease-acute or chronic; (2) the type of cell involved; myeloid (myelogenous), lymphoid (lymphogenous), or monocytic; and (3) the increase or non-increase in the number abnormal cells in the blood-leukemic or aleukemic (subleukemic). The murine leukemia model is widely accepted as being predictive of in vivo anti-leukemic activity.
  • the present invention includes a method of treating leukemia, including treating acute myeloid leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, a leukocythemic leukemia, basophylic leukemia, blast cell leukemia, bovine leukemia, chronic myelocytic leukemia, leukemia cutis, embryonal leukemia, eosinophilic leukemia, Gross' leukemia, hairy-cell leukemia, hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leuk
  • sarcoma generally refers to a tumor which is made up of a substance like the embryonic connective tissue and is generally composed of closely packed cells embedded in a fibrillar or homogeneous substance.
  • Sarcomas which can be treated with a combination of antineoplastic thiol-binding mitochondrial oxidant and an anticancer agent include a chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Abemethy's sarcoma, adipose sarcoma, liposarcoma, alveolar soft part sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma, chorio carcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrial sarcoma, stromal sarcoma, E
  • melanoma is taken to mean a tumor arising from the melanocytic system of the skin and other organs.
  • Melanomas which can be treated with a combination of antineoplastic thiol-binding mitochondrial oxidant and an anticancer agent include, for example, acral-lentiginous melanoma, amelanotic melanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma, Harding-Passey melanoma, juvenile melanoma, lentigo maligna melanoma, malignant melanoma, nodular melanoma, subungal melanoma, and superficial spreading melanoma.
  • carcinoma refers to a malignant new growth made up of epithelial cells tending to infiltrate the surrounding tissues and give rise to metastases.
  • exemplary carcinomas which can be treated with a combination of antineoplastic thiol-binding mitochondrial oxidant and an anticancer agent include, for example, acinar carcinoma, acinous carcinoma, adenocystic carcinoma, adenoid cystic carcinoma, carcinoma adenomatosum, carcinoma of adrenal cortex, alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma, carcinoma basocellulare, basaloid carcinoma, basosquamous cell carcinoma, bronchioalveolar carcinoma, bronchiolar carcinoma, bronchogenic carcinoma, cerebriform carcinoma, cholangiocellular carcinoma, chorionic carcinoma, colloid carcinoma, comedo carcinoma, corpus carcinoma, cribriform carcinoma, carcinoma en cuirasse, carcinoma cutaneum, cylindrical carcinoma, cylindrical cell carcinoma, duct carcinoma, carcinoma durum, embryonal carcinoma, encephaloid carcinoma,
  • Anti-cancer agent is used in accordance with its plain and ordinary meaning and refers to a composition (e.g. compound, drug, antagonist, inhibitor, modulator) having antineoplastic properties or the ability to inhibit the growth or proliferation of cells.
  • an anti-cancer agent is a chemotherapeutic.
  • An anti-cancer agent may be an agent approved by the FDA or similar regulatory agency of a country other than the USA, for treating cancer.
  • anti-cancer agents include, but are not limited to, MEK (e.g. MEK1, MEK2, or MEK1 and MEK2) inhibitors (e.g. XL518, CI-1040, PD035901, selumetinib/AZD6244, GSK1120212/trametinib, GDC-0973, ARRY-162, ARRY-300, AZD8330, PD0325901, U0126, PD98059, TAK-733, PD318088, AS703026, BAY 869766), alkylating agents (e.g., cyclophosphamide, ifosfamide, chlorambucil, busulfan, melphalan, mechlorethamine, uramustine, thiotepa, nitrosoureas, nitrogen mustards (e.g., mechloroethamine, cyclophosphamide, chlorambucil, meiphalan), ethy
  • TaxolTM i.e. paclitaxel
  • TaxotereTM compounds comprising the taxane skeleton, Erbulozole (i.e. R-55104), Dolastatin 10 (i.e. DLS-10 and NSC-376128), Mivobulin isethionate (i.e. as CI-980), Vincristine, NSC-639829, Discodermolide (i.e. as NVP-XX-A-296), ABT-751 (Abbott, i.e. E-7010), Altorhyrtins (e.g. Altorhyrtin A and Altorhyrtin C), Spongistatins (e.g.
  • Altorhyrtins e.g. Altorhyrtin A and Altorhyrtin C
  • Spongistatins e.g.
  • Epothilone E Epothilone F
  • Epothilone B N-oxide Epothilone A N-oxide
  • 16-aza-epothilone B Epothilone B
  • 21-aminoepothilone B i.e. BMS-310705
  • 21-hydroxyepothilone D i.e. Desoxyepothilone F and dEpoF
  • 26-fluoroepothilone i.e. NSC-654663
  • Soblidotin i.e. TZT-1027
  • LS-4559-P Pulacia, i.e.
  • LS-4577 LS-4578 (Pharmacia, i.e. LS-477-P), LS-4477 (Pharmacia), LS-4559 (Pharmacia), RPR-112378 (Aventis), Vincristine sulfate, DZ-3358 (Daiichi), FR-182877 (Fujisawa, i.e. WS-9885B), GS-164 (Takeda), GS-198 (Takeda), KAR-2 (Hungarian Academy of Sciences), BSF-223651 (BASF, i.e.
  • ILX-651 and LU-223651 SAH-49960 (Lilly/Novartis), SDZ-268970 (Lilly/Novartis), AM-97 (Armad/Kyowa Hakko), AM-132 (Armad), AM-138 (Armad/Kyowa Hakko), IDN-5005 (Indena), Cryptophycin 52 (i.e. LY-355703), AC-7739 (Ajinomoto, i.e. AVE-8063A and CS-39.HCl), AC-7700 (Ajinomoto, i.e.
  • T-900607 RPR-115781 (Aventis), Eleutherobins (such as Desmethyleleutherobin, Desaetyleleutherobin, Isoeleutherobin A, and Z-Eleutherobin), Caribaeoside, Caribaeolin, Halichondrin B, D-64131 (Asta Medica), D-68144 (Asta Medica), Diazonamide A, A-293620 (Abbott), NPI-2350 (Nereus), Taccalonolide A, TUB-245 (Aventis), A-259754 (Abbott), Diozostatin, ( ⁇ )-Phenylahistin (i.e.
  • NSCL-96F03-7 D-68838 (Asta Medica), D-68836 (Asta Medica), Myoseverin B, D-43411 (Zentaris, i.e. D-81862), A-289099 (Abbott), A-318315 (Abbott), HTI-286 (i.e.
  • SPA-10 trifluoroacetate salt
  • D-82317 Zentaris
  • D-82318 Zentaris
  • SC-12983 NCI
  • SSR-250411 SSR-250411 (Sanofi)
  • steroids e.g., dexamethasone
  • finasteride aromatase inhibitors
  • gonadotropin-releasing hormone agonists GnRH
  • adrenocorticosteroids e.g., prednisone
  • progestins e.g., hydroxyprogesterone caproate, megestrol acetate, medroxyprogesterone acetate
  • estrogens e.g., diethlystilbestrol, ethinyl estradiol
  • antiestrogen e.g., tamoxifen
  • androgens e.g.
  • gefitinib IressaTM
  • erlotinib TarcevaTM
  • cetuximab Erbitux
  • lapatinib TykerbTM
  • panitumumab VectibixTM
  • vandetanib CaprelsaTM
  • afatinib/BIBW2992 CI-1033/canertinib, neratinib/HKI-272, CP-724714, TAK-285, AST-1306, ARRY334543, ARRY-380, AG-1478, dacomitinib/PF299804, OSI-420/desmethyl erlotinib, AZD8931, AEE788, pelitinib/EKB-569, CUDC-101, WZ8040, WZ4002, WZ3146, AG-490, XL647, PD153035, BMS-599626), sorafenib, imatinib, sunitinib, dasatin
  • “Chemotherapeutic” or “chemotherapeutic agent” is used in accordance with its plain ordinary meaning and refers to a chemical composition or compound having antineoplastic properties or the ability to inhibit the growth or proliferation of cells.
  • Cancer model organism is an organism exhibiting a phenotype indicative of cancer, or the activity of cancer causing elements, within the organism.
  • the term cancer is defined above.
  • a wide variety of organisms may serve as cancer model organisms, and include for example, cancer cells and mammalian organisms such as rodents (e.g. mouse or rat) and primates (such as humans).
  • Cancer cell lines are widely understood by those skilled in the art as cells exhibiting phenotypes or genotypes similar to in vivo cancers. Cancer cell lines as used herein includes cell lines from animals (e.g. mice) and from humans.
  • the compound has formula (I):
  • A is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.
  • X is —C(R 4 ) ⁇ or —N ⁇ .
  • Y is a bond, —N(R 5 )—, —O—, or —S—.
  • L 1 is a bond, —C(O)—, —C(O)O—, —O—, —S—, —N(R 6 )—, —C(O)N(R 6 )—, —S(O) n6 —, —S(O)N(R 6 )—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene.
  • R 1 is halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 1A , —C(O)R 1A , —NR 1A R 1B , —C(O)OR 1A , —C(O)NR 1A R 1B , —NO 2 , —SR 1A , —S(O) n1 R 1A , —S(O) n1 OR 1A , —S(O) n1 NR 1A R 1B , —NHNR 1A R 1B , —ONR 1A R 1B , —NHC(O)NHNR 1A R 1B , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substitute
  • R 2 is halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 2A , —C(O)R 2A , —NR 2A R 2B , —C(O)OR 2A , —C(O)NR 2A R 2B , —NO 2 , —SR 2A , —S(O) n2 R 2A , —S(O) n2 OR 2A , —S(O) n2 NR 2A R 2B , —NHNR 2A R 2B , —ONR 2A R 2B , —NHC(O)NHNR 2A R 2B , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 3 is independently hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 4 is hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 4A , —C(O)R 4A , —NR 4A R 4B , —C(O)OR 4A , —C(O)NR 4A R 4B , —NO 2 , —SR 4A , —S(O) n4 R 4A , —S(O) n4 OR 4A , —S(O) n4 NR 4A R 4B , —NHNR 4A R 4B , —ONR 4A R 4B , —NHC(O)NHNR 4A R 4B , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 5 is hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 5A , —C(O)R 5A , —NR 5A R 5B , —C(O)OR 5A , —C(O)NR 5A R 5B , —NO 2 , —SR 5A , —S(O) n5 R 5A , —S(O) n5 OR 5A , —S(O) n5 NR 5A R 5B , —NHNR 5A R 5B , —ONR 5A R 5B , —NHC(O)NHNR 5A R 5B , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 6 is hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 6A , —C(O)R 6A , —NR 6A R 6B , —C(O)OR 6A , —C(O)NR 6A R 6B , —NO 2 , —SR 6A , —S(O)R 6A , —S(O) n6 OR 6A , —S(O) n6 NR 6A R 6B , —NHNR 6A R 6B , —ONR 6A R 6B , —NHC(O)NHNR 6A R 6B , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted
  • R 1A , R 1B , R 2A , R 2B , R 4A , R 4B , R 5A , R 5B , R 6A , and R 6B are independently hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 Cl, —S(O) 3 H, —S(O) 4 H, —S(O) 2 NH 2 , —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , —NHC(O)NH 2 , —NHS(O) 2 H, —NHC(O)H, —NHC(O)—OH, —NHOH, —OCF 3 , —OCHF 2 , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl
  • Ring A may be heterocycloalkyl, aryl, or heteroaryl. Ring A may be heterocycloalkyl or aryl. Ring A may be aryl or heteroaryl. Ring A may be heterocycloalkyl or heteroaryl.
  • Ring A may be cycloalkyl. Ring A may be a 3 to 10 membered cycloalkyl. Ring A may be 3 to 8 membered cycloalkyl. Ring A may be 3 to 6 membered cycloalkyl. Ring A may be cyclopropanyl. Ring A may be cyclopropenyl. Ring A may be cyclobutanyl. Ring A may be cyclobutenyl. Ring A may be cyclopentanyl. Ring A may be cyclopentenyl. Ring A may be cyclohexanyl. Ring A may be cyclohexenyl. Ring A may be cyclopentanyl, cyclopentenyl, cyclohexanyl, cyclohexenyl. Ring A may be cyclopentanyl, cyclopentenyl, cyclohexanyl, or cyclohexenyl.
  • Ring A may be heterocycloalkyl or heteroaryl. Ring A may be heterocycloalkyl. Ring A may be 3 to 10 membered heterocycloalkyl. Ring A may be 3 to 8 membered heterocycloalkyl. Ring A may be 3 to 6 membered heterocycloalkyl. Ring A may be 3 membered heterocycloalkyl. Ring A may be 4 membered heterocycloalkyl.
  • Ring A may be aziridinyl, azirinyl, oxiranyl, oxirenyl, thiiranyl, thiirenyl, azetidinyl, azetyl, oxetanyl, oxetyl, thietanyl, or thietyl.
  • Ring A may be 5 membered heterocycloalkyl. Ring A may be 6 membered heterocycloalkyl. Ring A may be tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopyranyl, pyranyl, thianyl, dioxanyl, piperidinyl, piperazinyl, oxathianyl, morpholinyl, trioxanyl, pyrrolinyl, pyrrolidinyl, pyrrolidonyl, pyrrolidionyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, isoxazolinyl, isoxazolidinyl, oxazolinyl, oxazolidinyl, oxazolidinonyl, or thiazolidinyl
  • Ring A may be aryl. Ring A may be 5 to 10 membered aryl. Ring A may be 5 or 6 membered aryl. Ring A may be 5 membered aryl. Ring A may be 6 membered aryl. Ring A may be phenylene. Ring A may be heteroaryl. Ring A may be 5 to 10 membered heteroaryl. Ring A may be 5 or 6 membered heteroaryl. Ring A may be 5 membered heteroaryl. Ring A may be 6 membered heteroaryl.
  • Ring A may be pyrroyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, pyrazoyl, furyl, thienyl, triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl.
  • Ring A may be a 5,6-, 6,5-, or 6,6-fused ring aryl or 5,6-, 6,5-, or 6,6-fused ring heteroaryl as described herein. Ring A may be a 5,6-fused ring aryl. Ring A may be a 5,6-fused ring heteroaryl. Ring A may be a 6,5-fused ring aryl. Ring A may be a 6,5-fused ring heteroaryl. Ring A may be 6,6-fused ring aryl. Ring A may be a 6,6-fused ring heteroaryl.
  • Ring A may indenyl, indolyl, isoindolyl, indolizinyl, purinyl, benzothiazolyl, benzoxazoyl, benzoimidazoyl, benzofuranyl, isobenzofuranyl indazolyl, pyrrollopyridinyl, pyrrollopyrimidinyl, pyrazolopyridinyl, imidazopyridinyl, benzotriazolyl, benzothiophenyl, quinolyl, quinolinyl, isoquinolyl, naphthalenyl, cinnolinyl, phthalazinyl, isoquinolinyl, quinoxalinyl, or quinazolinyl.
  • the symbol m2 may be 0.
  • the symbol m2 may be 1.
  • the symbol m2 may be 1, 2, 3, 4, 5, or 6.
  • the symbol m2 may be 1, 2, or 3.
  • the symbols n1, n2, n4, n5, and n6 may independently be 1.
  • the symbols n1, n2, n4, n5, and n6 may independently be 2.
  • the symbols n1, n2, n4, n5, and n6 may independently be 3.
  • R 1 may be halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 1A , —C(O)R 1A , —NR 1A R 1B , —C(O)OR 1A , —C(O)NR 1A R 1B , —NO 2 , —SR 1A , —S(O) n1 R 1A , —S(O) n1 OR 1A , —S(O) n1 NR 1A R 1B , —NHNR 1A R 1B , —ONR 1A R 1B , —NHC(O)NHNR 1A R 1B .
  • R 1 may be halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 1A , —C(O)R 1A , —NR 1A R 1B , —C(O)OR 1A , —C(O)NR 1A R 1B , —NO 2 , —SR 1A .
  • R 1 may be halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 1A , —NH 2 , —C(O)OH, —C(O)NH 2 , —NO 2 , or —SH.
  • R 1 may be halogen, —CF 3 , —NO 2 , —NH 2 , —OR 1A .
  • R 1 may be halogen.
  • R 1 may be F.
  • R 1 may be Cl.
  • R 1 may be Br.
  • R 1 may be I.
  • R 1 may be —CF 3 .
  • R 1 may be —NH 2 .
  • R 1 may be —OR 1A , where R 1A is as defined herein.
  • R 1 may be —OR 1A where R 1A is substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, or substituted or unsubstituted aryl.
  • R 1 may be —OR 1A where R 1A is substituted or unsubstituted alkyl.
  • R 1 may be —OR 1A where R 1A is unsubstituted alkyl.
  • R 1 may be —OR 1A where R 1A is unsubstituted C 1 -C 5 alkyl.
  • R 1 may be —OR 1A where R 1A is unsubstituted C 1 -C 3 alkyl.
  • R 1 may be —OR 1A where R 1A is methyl (e.g. —OCH 3 ).
  • R 1 may be halogen, —CF 3 , —NO 2 , —NH 2 , —OR 1A , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • R 1 may be substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • R 1 may be substituted or unsubstituted alkyl.
  • R 1 may be substituted alkyl.
  • R 1 may be unsubstituted alkyl.
  • R 1 may be substituted or unsubstituted C 1 -C 20 alkyl.
  • R 1 may be substituted C 1 -C 20 alkyl.
  • R 1 may be substituted or unsubstituted C 1 -C 10 alkyl.
  • R 1 may be substituted C 1 -C 10 alkyl.
  • R 1 may be unsubstituted C 1 -C 10 alkyl.
  • R 1 may be substituted or unsubstituted C 1 -C 5 alkyl.
  • R 1 may be substituted C 1 -C 5 alkyl.
  • R 1 may be unsubstituted C 1 -C 5 alkyl.
  • R 1 may be substituted or unsubstituted C 1 -C 3 alkyl.
  • R 1 may be substituted C 1 -C 3 alkyl.
  • R 1 may be unsubstituted C 1 -C 3 alkyl.
  • R 1 may be methyl.
  • R 1 may be ethyl.
  • R 1 may be propyl.
  • R 1 may be R 10 -substituted or unsubstituted alkyl.
  • R 1 may be R 10 -substituted alkyl.
  • R 1 may be R 10 -substituted or unsubstituted C 1 -C 20 alkyl.
  • R 1 may be substituted C 1 -C 20 alkyl.
  • R 1 may be R 10 -substituted or unsubstituted C 1 -C 10 alkyl.
  • R 1 may be R 10 -substituted C 1 -C 10 alkyl.
  • R 1 may be R 10 -substituted or unsubstituted C 1 -C 5 alkyl.
  • R 1 may be R 10 -substituted C 1 -C 5 alkyl.
  • R 1 may be R 10 -substituted or unsubstituted C 1 -C 3 alkyl.
  • R 1 may be R 10 -substituted C 1 -C 3 alkyl.
  • R 1 may be substituted or unsubstituted 2 to 20 membered heteroalkyl.
  • R 1 may be substituted 2 to 20 membered heteroalkyl.
  • R 1 may be unsubstituted 2 to 20 membered heteroalkyl.
  • R 1 may be substituted or unsubstituted 2 to 10 membered heteroalkyl.
  • R 1 may be substituted 2 to 10 membered heteroalkyl.
  • R 1 may be unsubstituted 2 to 10 membered heteroalkyl.
  • R 1 may be substituted or unsubstituted 2 to 6 membered heteroalkyl.
  • R 1 may be substituted 2 to 6 membered heteroalkyl.
  • R 1 may be unsubstituted 2 to 6 membered heteroalkyl.
  • R 1 may be substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 1 may be substituted 2 to 5 membered heteroalkyl.
  • R 1 may be substitute
  • R 1 may be R 10 -substituted or unsubstituted 2 to 20 membered heteroalkyl.
  • R 1 may be R 10 -substituted 2 to 20 membered heteroalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 2 to 10 membered heteroalkyl.
  • R 1 may be R 10 -substituted 2 to 10 membered heteroalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 2 to 6 membered heteroalkyl.
  • R 1 may be R 10 -substituted 2 to 6 membered heteroalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 1 may be R 10 -substituted 2 to 5 membered heteroalkyl.
  • R 1 may be substituted or unsubstituted 3 to 10 membered cycloalkyl.
  • R 1 may be substituted 3 to 10 membered cycloalkyl.
  • R 1 may be unsubstituted 3 to 10 membered cycloalkyl.
  • R 1 may be substituted or unsubstituted 3 to 6 membered cycloalkyl.
  • R 1 may be substituted 3 to 6 membered cycloalkyl.
  • R 1 may be unsubstituted 3 to 6 membered cycloalkyl.
  • R 1 may be substituted or unsubstituted 3 to 5 membered cycloalkyl.
  • R 1 may be substituted 3 to 5 membered cycloalkyl.
  • R 1 may be unsubstituted 3 to 5 membered cycloalkyl.
  • R 1 may be substituted or unsubstituted 3 membered cycloalkyl.
  • R 1 may be substituted 3 membered cycloalkyl.
  • R 1 may be unsubstituted 3 membered cycloalkyl.
  • R 1 may be substituted or unsubstituted 4 membered cycloalkyl.
  • R 1 may be substituted 4 membered cycloalkyl.
  • R 1 may be unsubstituted 4 membered cycloalkyl.
  • R 1 may be substituted or unsubstituted 5 membered cycloalkyl.
  • R 1 may be substituted 5 membered cycloalkyl.
  • R 1 may be unsubstituted 5 membered cycloalkyl.
  • R 1 may be substituted or unsubstituted 6 membered cycloalkyl.
  • R 1 may be substituted 6 membered cycloalkyl.
  • R 1 may be unsubstituted 6 membered cycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 3 to 10 membered cycloalkyl.
  • R 1 may be R 10 -substituted 3 to 10 membered cycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 3 to 6 membered cycloalkyl.
  • R 1 may be R 10 -substituted 3 to 6 membered cycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 3 to 5 membered cycloalkyl.
  • R 1 may be R 10 -substituted 3 to 5 membered cycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 3 membered cycloalkyl.
  • R 1 may be R 10 -substituted 3 membered cycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 4 membered cycloalkyl.
  • R 1 may be R 10 -substituted 4 membered cycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 5 membered cycloalkyl.
  • R 1 may be R 10 -substituted 5 membered cycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 6 membered cycloalkyl.
  • R 1 may be R 10 -substituted 6 membered cycloalkyl.
  • R 1 may be substituted or unsubstituted 3 to 10 membered heterocycloalkyl.
  • R 1 may be substituted 3 to 10 membered heterocycloalkyl.
  • R 1 may be unsubstituted 3 to 10 membered heterocycloalkyl.
  • R 1 may be substituted or unsubstituted 3 to 6 membered heterocycloalkyl.
  • R 1 may be substituted 3 to 6 membered heterocycloalkyl.
  • R 1 may be unsubstituted 3 to 6 membered heterocycloalkyl.
  • R 1 may be substituted or unsubstituted 3 to 5 membered heterocycloalkyl.
  • R 1 may be substituted 3 to 5 membered heterocycloalkyl.
  • R 1 may be unsubstituted 3 to 5 membered heterocycloalkyl.
  • R 1 may be substituted or unsubstituted 3 membered heterocycloalkyl.
  • R 1 may be substituted 3 membered heterocycloalkyl.
  • R 1 may be unsubstituted 3 membered heterocycloalkyl.
  • R 1 may be substituted or unsubstituted 4 membered heterocycloalkyl.
  • R 1 may be substituted 4 membered heterocycloalkyl.
  • R 1 may be unsubstituted 4 membered heterocycloalkyl.
  • R 1 may be substituted or unsubstituted 5 membered heterocycloalkyl.
  • R 1 may be substituted 5 membered heterocycloalkyl.
  • R 1 may be unsubstituted 5 membered heterocycloalkyl.
  • R 1 may be substituted or unsubstituted 6 membered heterocycloalkyl.
  • R 1 may be substituted 6 membered heterocycloalkyl.
  • R 1 may be unsubstituted 6 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 3 to 10 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted 3 to 10 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 3 to 6 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted 3 to 6 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 3 to 5 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted 3 to 5 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 3 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted 3 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 4 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted 4 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 5 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted 5 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted or unsubstituted 6 membered heterocycloalkyl.
  • R 1 may be R 10 -substituted 6 membered heterocycloalkyl.
  • R 1 may be substituted or unsubstituted aryl.
  • R 1 may be substituted aryl.
  • R 1 may be unsubstituted aryl.
  • R 1 may be substituted or unsubstituted 5 to 10 membered aryl.
  • R 1 may be substituted 5 to 10 membered aryl.
  • R 1 may be unsubstituted 5 to 10 membered aryl.
  • R 1 may be substituted or unsubstituted 5 or 6 membered aryl.
  • R 1 may be substituted 5 or 6 membered aryl.
  • R 1 may be unsubstituted 5 or 6 membered aryl.
  • R 1 may be substituted or unsubstituted 5 membered aryl.
  • R 1 may be substituted 5 membered aryl.
  • R 1 may be unsubstituted 5 membered aryl.
  • R 1 may be substituted or unsubstituted 6 membered aryl.
  • R 1 may be substituted 6 membered aryl.
  • R 1 may be unsubstituted 6 membered aryl.
  • R 1 may be R 10 -substituted or unsubstituted aryl.
  • R 1 may be R 10 -substituted aryl.
  • R 1 may be R 10 -substituted or unsubstituted 5 to 10 membered aryl.
  • R 1 may be R 10 -substituted 5 to 10 membered aryl.
  • R 1 may be R 10 -substituted or unsubstituted 5 or 6 membered aryl.
  • R 1 may be R 10 -substituted 5 or 6 membered aryl.
  • R 1 may be R 10 -substituted or unsubstituted 5 membered aryl.
  • R 1 may be R 10 -substituted 5 membered aryl.
  • R 1 may be R 10 -substituted or unsubstituted 6 membered aryl.
  • R 1 may be R 10 -substituted 6 membered aryl.
  • R 1 may be substituted or unsubstituted heteroaryl.
  • R 1 may be substituted heteroaryl.
  • R 1 may be unsubstituted heteroaryl.
  • R 1 may be substituted or unsubstituted 5 to 10 membered heteroaryl.
  • R 1 may be substituted 5 to 10 membered heteroaryl.
  • R 1 may be unsubstituted 5 to 10 membered heteroaryl.
  • R 1 may be substituted or unsubstituted 5 or 6 membered heteroaryl.
  • R 1 may be substituted 5 or 6 membered heteroaryl.
  • R 1 may be unsubstituted 5 or 6 membered heteroaryl.
  • R 1 may be substituted or unsubstituted 5 membered heteroaryl.
  • R 1 may be substituted 5 membered heteroaryl.
  • R 1 may be unsubstituted 5 membered heteroaryl.
  • R 1 may be substituted or unsubstituted 6 membered heteroaryl.
  • R 1 may be substituted 6 membered heteroaryl.
  • R 1 may be unsubstituted 6 membered heteroaryl.
  • R 1 may be R 10 -substituted or unsubstituted heteroaryl.
  • R 1 may be R 10 -substituted heteroaryl.
  • R 1 may be R 10 -substituted or unsubstituted 5 to 10 membered heteroaryl.
  • R 1 may be R 10 -substituted 5 to 10 membered heteroaryl.
  • R 1 may be R 10 -substituted or unsubstituted 5 or 6 membered heteroaryl.
  • R 1 may be R 10 -substituted 5 or 6 membered heteroaryl.
  • R 1 may be R 10 -substituted or unsubstituted 5 membered heteroaryl.
  • R 1 may be R 10 -substituted 5 membered heteroaryl.
  • R 1 may be R 10 -substituted or unsubstituted 6 membered heteroaryl.
  • R 1 may be R 10 -substituted 6 membered heteroaryl.
  • R 10 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , R 11 -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 11 -substituted or unsubstituted (e.g.
  • R 11 -substituted or unsubstituted e.g. C 3 -C 8 cycloalkyl
  • R 11 -substituted or unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl e.g. 1,3-butaned
  • R 11 -substituted or unsubstituted aryl e.g. phenyl or naphthyl
  • R 1 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 10 may be hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, substituted or unsubstituted (e.g.
  • heteroalkyl substituted or unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • substituted or unsubstituted e.g. C 3 -C 8
  • substituted or unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl substituted or unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 10 may be hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , unsubstituted (e.g. C 1 -C 5 ) alkyl, unsubstituted (e.g.
  • heteroalkyl 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • unsubstituted e.g. C 3 -C 8
  • unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • R 11 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , R 12 -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 12 -substituted or unsubstituted (e.g.
  • R 12 -substituted or unsubstituted e.g. C 3 -C 8 cycloalkyl
  • R 12 -substituted or unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl e.g. 1,3-butaned
  • R 12 -substituted or unsubstituted aryl e.g. phenyl or naphthyl
  • R 12 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 12 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , R 13 -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 13 -substituted or unsubstituted (e.g.
  • R 13 -substituted or unsubstituted e.g. C 3 -C 8 cycloalkyl
  • R 13 -substituted or unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl e.g. 1,3-butaned
  • R 13 -substituted or unsubstituted aryl e.g. phenyl or naphthyl
  • R 13 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 13 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , unsubstituted (e.g. C 1 -C 5 ) alkyl, unsubstituted (e.g.
  • heteroalkyl 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • unsubstituted e.g. C 3 -C 8
  • unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • the symbol m1 may be 0, 1, 2, or 3.
  • the symbol m1 may be 0.
  • the symbol m1 may be 1.
  • the symbol m1 may be 2.
  • the symbol m1 may be 3.
  • the symbol m1 may be 1, 2, or 3.
  • R 1 is halogen
  • the symbol m1 may be 1.
  • R 1 is —CF 3
  • R 1 may independently be halogen or —CF 3 and the symbol m1 is 2 or 3.
  • R 1 is —OR 1A
  • the symbol m1 may be 1, 2, or 3.
  • R 1 is —OCH 3
  • the symbol m1 may be 1, 2, or 3.
  • When R 1 is —OCH 3 the symbol m1 may be 1.
  • R 1 is —OCH 3
  • the symbol m1 may be 2.
  • R 1 is —OCH 3
  • the symbol m1 may be 3.
  • R 1A and R 1B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, —ONH 2 , —NHC(O)NHNH 2 , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • R 1A and R 1B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, —ONH 2 , —NHC(O)NHNH 2 , R 10 -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 10 -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, R 10 -substituted or unsubstituted (e.g.
  • C 3 -C 8 cycloalkyl, R 10 -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 10 -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 10 -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • aryl e.g. phenyl or naphthyl
  • R 10 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 1A and R 1B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, or —ONH 2 .
  • R 1A and R 1B may independently be substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • R 1A and R 1B may independently be R 1C -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 1C -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, R 1C -substituted or unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, R 1C -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 1C -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 1C -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • R 1C -substituted or unsubstituted e.g. C 1 -
  • R 1C is hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 Cl, —S(O) 3 H, —S(O) 4 H, —S(O) 2 NH 2 , —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , —NHC(O)NH 2 , —NHS(O) 2 H, —NHC(O)H, —NHC(O)—OH, —NHOH, —OCF 3 , —OCHF 2 , unsubstituted (e.g.
  • C 1 -C 5 alkyl unsubstituted (e.g. 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • R 2 may be halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 2A , —C(O)R 2A , —NR 2A R 2B , —C(O)OR 2A , —C(O)NR 2A R 2B , —NO 2 , —SR 2A , —S(O) n2 R 2A , —S(O) n2 OR 2A , —S(O) n2 NR 2A R 2B , —NHNR 2A R 2B , —ONR 2A R 2B , or —NHC(O)NHNR 2A R 2B .
  • R 2 may be halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 2A , —NR 2A R 2B , —NO 2 , or —SR 2A .
  • R 2 may be halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 2A , —C(O)R 2A , —NR 2A R 2B , —C(O)OH, —NO 2 , or —SH.
  • R 2 may be halogen, —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —OR 2A , —NH 2 , —N(CH 3 ) 2 —NO 2 .
  • R 2 may be halogen, —CF 3 , —OR 2A , or —NO 2 .
  • R 2 may be halogen.
  • R 2 may be —CF 3 .
  • R 2 may be —OR 2A .
  • R 2 may be —NO 2 .
  • R 2 may be —OR 2A , where R 2A is substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, or substituted or unsubstituted aryl.
  • R 2 may be —OR 2A , where R 2A is substituted or unsubstituted alkyl.
  • R 2 may be —OR 2A , where R 2A is substituted or unsubstituted C 1 -C 5 alkyl.
  • R 2 may be —OR 2A , where R 2A is substituted or unsubstituted C 1 -C 3 alkyl.
  • R 2 may be —OR 2A , where R 2A is methyl.
  • R 2 may be halogen, —CF 3 , —OR 2A , —NO 2 , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 2 may be substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 2 may be substituted or unsubstituted C 1 -C 5 alkyl.
  • R 2 may be substituted C 1 -C 5 alkyl.
  • R 2 may be unsubstituted C 1 -C 5 alkyl.
  • R 2 may be substituted or unsubstituted C 1 -C 3 alkyl.
  • R 2 may be substituted C 1 -C 3 alkyl.
  • R 2 may be unsubstituted C 1 -C 3 alkyl.
  • R 2 may be methyl.
  • R 2 may be ethyl.
  • R 2 may be propyl.
  • R 2 may be R 20 -substituted or unsubstituted C 1 -C 5 alkyl.
  • R 2 may be R 20 -substituted or unsubstituted C 1 -C 5 alkyl.
  • R 2 may be R 20 -substituted C 1 -C 5 alkyl.
  • R 2 may be R 20 -substituted or unsubstituted C 1 -C 3 alkyl.
  • R 2 may be R 20 -substituted C 1 -C 3 alkyl.
  • R 2 may be substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 2 may be substituted 2 to 5 membered heteroalkyl.
  • R 2 may be unsubstituted 2 to 5 membered heteroalkyl.
  • R 2 may be R 20 -substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 2 may be R 20 -substituted 2 to 5 membered heteroalkyl.
  • R 20 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , unsubstituted (e.g. C 1 -C 5 ) alkyl, unsubstituted (e.g.
  • heteroalkyl 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • unsubstituted e.g. C 3 -C 8
  • unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • R 2A and R 2B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, —ONH 2 , —NHC(O)NHNH 2 , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • R 2A and R 2B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, —ONH 2 , —NHC(O)NHNH 2 , R 20 -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 20 -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, R 20 -substituted or unsubstituted (e.g.
  • C 3 -C 8 cycloalkyl, R 20 -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 20 -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 20 -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • aryl e.g. phenyl or naphthyl
  • R 20 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 2A and R 2B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, or —ONH 2 .
  • R 2A and R 2B may independently be substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • R 2A and R 2B may independently be R 2C -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 2C -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, R 2C -substituted or unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, R 2C -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 2C -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 2C -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • R 2C -substituted or unsubstituted e.g. C 1 -
  • R 2C is hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 Cl, —S(O) 3 H, —S(O) 4 H, —S(O) 2 NH 2 , —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , —NHC(O)NH 2 , —NHS(O) 2 H, —NHC(O)H, —NHC(O)—OH, —NHOH, —OCF 3 , —OCHF 2 , unsubstituted (e.g.
  • C 1 -C 5 alkyl unsubstituted (e.g. 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • R 3 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , or —NHC(O)NHNH 2 .
  • R 3 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , or —ONH 2 .
  • R 3 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , or —SH.
  • R 3 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OCH 3 , —OCH 2 CH 3 , —NH 2 , or —NO 2 .
  • R 3 may be hydrogen, halogen, or —OR 3A .
  • R 3 may be hydrogen, halogen, —CF 3 , —OCH 3 , —OCH 2 CH 3 , —NH 2 , or —NO 2 .
  • R 3 may be halogen.
  • R 3 may be hydrogen.
  • R 3 may be —CF 3 .
  • R 3 may be —OCH 3 .
  • R 3 may be —OCH 2 CH 3 .
  • R 3 may be —NH 2 .
  • R 3 may be —NO 2 .
  • R 3 may be substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 3 may be substituted or unsubstituted C 1 -C 5 alkyl.
  • R 3 may be substituted C 1 -C 5 alkyl.
  • R 3 may be unsubstituted C 1 -C 5 alkyl.
  • R 3 may be substituted or unsubstituted C 1 -C 3 alkyl.
  • R 3 may be substituted C 1 -C 3 alkyl.
  • R 3 may be unsubstituted C 1 -C 3 alkyl.
  • R 3 may be methyl.
  • R 3 may be ethyl.
  • R 3 may be propyl.
  • R 3 may be R 30 -substituted or unsubstituted C 1 -C 5 alkyl.
  • R 3 may be R 30 -substituted or unsubstituted C 1 -C 5 alkyl.
  • R 3 may be R 30 -substituted C 1 -C 5 alkyl.
  • R 3 may be R 30 -substituted or unsubstituted C 1 -C 3 alkyl.
  • R 3 may be R 30 -substituted C 1 -C 3 alkyl.
  • R 3 may be substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 3 may be substituted 2 to 5 membered heteroalkyl.
  • R 3 may be unsubstituted 2 to 5 membered heteroalkyl.
  • R 3 may be R 30 -substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 3 may be R 30 -substituted 2 to 5 membered heteroalkyl.
  • R 30 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , unsubstituted (e.g. C 1 -C 5 ) alkyl, unsubstituted (e.g.
  • heteroalkyl 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • unsubstituted e.g. C 3 -C 8
  • unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • the symbol m3 may be 1.
  • the symbol m3 may be 2.
  • R 3 is halogen
  • the symbol m3 may be 1.
  • R 3 is —CF 3
  • the symbol m3 may be 1.
  • R 3 is —OCH 3 or —OCH 2 CH 3
  • the symbol m3 may be 1.
  • R 3 is —OCH 3 or —OCH 2 CH 3
  • the symbol m3 may be 2.
  • R 3 may independently be halogen and —OCH 3 and the symbol m3 is 2.
  • X may be —C(R 4 ) ⁇ .
  • X may be —N ⁇ .
  • R 4 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 4A , —C(O)R 4A , —NR 4A R 4B , —C(O)OR 4A , —C(O)NR 4A R 4B , —NO 2 , —SR 4A , —S(O) n4 R 4A , —S(O) n4 OR 4A , —S(O) n4 NR 4A R 4B , —NHNR 4A R 4B , —ONR 4A R 4B , or —NHC(O)NHNR 4A R 4 .
  • R 4 may be hydrogen, halogen, —OR 4A , or —C(O)R 4A .
  • R 4 may be hydrogen or halogen.
  • R 4 may be hydrogen
  • R 4 may be substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 4 may be substituted or unsubstituted C 1 -C 5 alkyl.
  • R 4 may be substituted C 1 -C 5 alkyl.
  • R 4 may be unsubstituted C 1 -C 5 alkyl.
  • R 4 may be substituted or unsubstituted C 1 -C 3 alkyl.
  • R 4 may be substituted C 1 -C 3 alkyl.
  • R 4 may be unsubstituted C 1 -C 3 alkyl.
  • R 4 may be methyl.
  • R 4 may be ethyl.
  • R 4 may be propyl.
  • R 4 may be R 40 -substituted or unsubstituted C 1 -C 5 alkyl.
  • R 4 may be R 40 -substituted or unsubstituted C 1 -C 5 alkyl.
  • R 4 may be R 40 -substituted C 1 -C 5 alkyl.
  • R 4 may be R 40 -substituted or unsubstituted C 1 -C 3 alkyl.
  • R 4 may be R 40 -substituted C 1 -C 3 alkyl.
  • R 4 may be substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 4 may be substituted 2 to 5 membered heteroalkyl.
  • R 4 may be unsubstituted 2 to 5 membered heteroalkyl.
  • R 4 may be R 40 -substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 4 may be R 40 -substituted 2 to 5 membered heteroalkyl.
  • R 40 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , unsubstituted (e.g. C 1 -C 5 ) alkyl, unsubstituted (e.g.
  • heteroalkyl 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • unsubstituted e.g. C 3 -C 8
  • unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • R 4A and R 4B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, —ONH 2 , —NHC(O)NHNH 2 , R 40 -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 40 -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, R 40 -substituted or unsubstituted (e.g.
  • R 40 C 3 -C 8 ) cycloalkyl, R 40 -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 40 -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 40 -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • aryl e.g. phenyl or naphthyl
  • R 40 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 4A and R 4B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, or —ONH 2 .
  • R 4A and R 4B may independently be substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • R 4A and R 4B may independently be R 4C -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 4C -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, R 4C -substituted or unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, R 4C -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 4C -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 4C -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • R 4C -substituted or unsubstituted e.g. C 1 -
  • R 4C is hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 Cl, —S(O) 3 H, —S(O) 4 H, —S(O) 2 NH 2 , —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , —NHC(O)NH 2 , —NHS(O) 2 H, —NHC(O)H, —NHC(O)—OH, —NHOH, —OCF 3 , —OCHF 2 , unsubstituted (e.g.
  • C 1 -C 5 alkyl unsubstituted (e.g. 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • Y may be a bond or —N(R 5 )—. Y may be a bond or —O—. Y may be a bond or —S—. Y may be a bond, —N(R 5 )—, or —S—. Y may be a bond. Y may be —O—. Y may be —S—. Y may be —N(R 5 )—.
  • R 5 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 5A , —C(O)R 5A , —NR 5A R 5B , —C(O)OR 5A , —C(O)NR 5A R 5B , —NO 2 , —SR 5A , —S(O) n5 R 5A , —S(O) n5 OR 5A , —S(O) n5 NR 5A R 5B , —NHNR 5A R 5B , —ONR 5A R 5B , or —NHC(O)NHNR 5A R 5B .
  • R 5 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 5A , —NR 5A R 5B , —C(O)OR 5A , —C(O)NR 5A R 5B , —NO 2 , or —SR 5A .
  • R 5 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 5A , —NR 5A R 5B , —C(O)OR 5A , —NO 2 , or —SR 5A .
  • R 5 may be hydrogen, halogen, —CF 3 , —CN, —OR 5A , —NR 5A R 5B , —NO 2 , or —SR 5A .
  • R 5 may be hydrogen, halogen, —CF 3 , —CN, —OR 5A , —NH 2 , —NO 2 , or —SH.
  • R 5 may be hydrogen.
  • R 5 may be halogen.
  • R 5 may be —CF 3 .
  • R 5 may be —CN.
  • R 5 may be —OR 5A .
  • R 5 may be —NR 5A R 5B .
  • R 5 may be, —NO 2 .
  • R 5 may be —SR 5A .
  • R 5 may be substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 5 may be substituted or unsubstituted C 1 -C 5 alkyl.
  • R 5 may be substituted C 1 -C 5 alkyl.
  • R 5 may be unsubstituted C 1 -C 5 alkyl.
  • R 5 may be substituted or unsubstituted C 1 -C 3 alkyl.
  • R 5 may be substituted C 1 -C 3 alkyl.
  • R 5 may be unsubstituted C 1 -C 3 alkyl.
  • R 5 may be methyl.
  • R 5 may be ethyl.
  • R 5 may be propyl.
  • R 5 may be R 50 -substituted or unsubstituted C 1 -C 5 alkyl.
  • R 5 may be R 50 -substituted or unsubstituted C 1 -C 5 alkyl.
  • R 5 may be R 50 -substituted C 1 -C 5 alkyl.
  • R 5 may be R 50 -substituted or unsubstituted C 1 -C 3 alkyl.
  • R 5 may be R 50 -substituted C 1 -C 3 alkyl.
  • R 5 may be substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 5 may be substituted 2 to 5 membered heteroalkyl.
  • R 5 may be unsubstituted 2 to 5 membered heteroalkyl.
  • R 5 may be R 50 -substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 5 may be R 5 -substituted 2 to 5 membered heteroalkyl.
  • R 50 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , unsubstituted (e.g. C 1 -C 5 ) alkyl, unsubstituted (e.g.
  • heteroalkyl 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • unsubstituted e.g. C 3 -C 8
  • unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • R 5A and R 5B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, or —ONH 2 .
  • R 5A and R 5B may independently be substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • R 5A and R 5B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, —ONH 2 , —NHC(O)NHNH 2 , R 50 -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 50 -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, R 50 -substituted or unsubstituted (e.g.
  • C 3 -C 8 cycloalkyl, R 50 -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 5 -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 50 -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • aryl e.g. phenyl or naphthyl
  • R 50 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 5A and R 5B may independently be R 5C -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 5C -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, R 5C -substituted or unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, R 5C -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 5C -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 5C -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • R 5C -substituted or unsubstituted e.g. C 1 -
  • R 5C is hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 Cl, —S(O) 3 H, —S(O) 4 H, —S(O) 2 NH 2 , —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , —NHC(O)NH 2 , —NHS(O) 2 H, —NHC(O)H, —NHC(O)—OH, —NHOH, —OCF 3 , —OCHF 2 , unsubstituted (e.g.
  • C 1 -C 5 alkyl unsubstituted (e.g. 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • L 1 may be a bond, —C(O)—, —O—, —S—, —N(R 6 )—, —C(O)N(R 6 )—, —S(O) n6 —, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene.
  • L 1 may be a bond, —C(O)—, —O—, —S—, —NH—, —C(O)NH—, —S(O) 2 —, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene.
  • L 1 may be a bond, —C(O)—, —O—, —NH—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene.
  • L 1 may be a bond, —C(O)—, —O—, —S—, —N(R 6 )—, —C(O)N(R 6 )—, or —S(O) n6 —.
  • L 1 may be a bond, —C(O)—, —O—, —S—, —NH—, —C(O)NH—, or —S(O) 2 —.
  • L 1 may be a bond.
  • L 1 may be —C(O).
  • L 1 may be —O—.
  • L 1 may be —S—.
  • L 1 may —N(R 6 )—.
  • L 1 may be —C(O)N(R 6 ).
  • L 1 may be —S(O) n6 —.
  • L 1 may be substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene.
  • L 1 may be substituted or unsubstituted alkylene.
  • L 1 may be substituted alkylene.
  • L 1 may be unsubstituted alkylene.
  • L 1 may be substituted or unsubstituted C 1 -C 20 alkylene.
  • L 1 may be substituted C 1 -C 20 alkylene.
  • L 1 may be substituted or unsubstituted C 1 -C 10 alkylene.
  • L 1 may be substituted C 1 -C 10 alkylene.
  • L 1 may be unsubstituted C 1 -C 10 alkylene.
  • L 1 may be substituted or unsubstituted C 1 -C 5 alkylene.
  • L 1 may be substituted C 1 -C 5 alkylene.
  • L 1 may be unsubstituted C 1 -C 5 alkylene.
  • L 1 may be R 14 -substituted or unsubstituted alkylene.
  • L 1 may be R 14 -substituted alkylene.
  • L 1 may be R 14 -substituted or unsubstituted C 1 -C 20 alkylene.
  • L 1 may be R 14 -substituted C 1 -C 20 alkylene.
  • L 1 may be R 14 -substituted or unsubstituted C 1 -C 10 alkylene.
  • L 1 may be R 14 -substituted C 1 -C 10 alkylene.
  • L 1 may be R 14 -substituted or unsubstituted C 1 -C 5 alkylene.
  • L 1 may be R 14 -substituted C 1 -C 5 alkylene.
  • L 1 may be substituted or unsubstituted heteroalkylene.
  • L 1 may be substituted heteroalkylene.
  • L 1 may be unsubstituted heteroalkylene.
  • L 1 may be substituted or unsubstituted 2 to 20 membered heteroalkylene.
  • L 1 may be substituted 2 to 20 membered heteroalkylene.
  • L 1 may be unsubstituted 2 to 20 membered heteroalkylene.
  • L 1 may be substituted or unsubstituted 2 to 10 membered heteroalkylene.
  • L 1 may be substituted 2 to 10 membered heteroalkylene.
  • L 1 may be unsubstituted 2 to 10 membered heteroalkylene.
  • L 1 may be substituted or unsubstituted 2 to 5 membered heteroalkylene.
  • L 1 may be substituted 2 to 5 membered heteroalkylene.
  • L 1 may be unsubstituted 2 to 5 membered heteroalkylene.
  • L 1 may be R 14 -substituted or unsubstituted heteroalkylene.
  • L 1 may be R 14 -substituted heteroalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 2 to 20 membered heteroalkylene.
  • L 1 may be R 14 -substituted 2 to 20 membered heteroalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 2 to 10 membered heteroalkylene.
  • L 1 may be R 14 -substituted 2 to 10 membered heteroalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 2 to 5 membered heteroalkylene.
  • L 1 may be R 14 -substituted 2 to 5 membered heteroalkylene.
  • L 1 may be substituted or unsubstituted cycloalkylene.
  • L 1 may be substituted cycloalkylene.
  • L 1 may be unsubstituted cycloalkylene.
  • L 1 may be substituted or unsubstituted 3 to 10 membered cycloalkylene.
  • L 1 may be substituted 3 to 10 membered cycloalkylene.
  • L 1 may be unsubstituted 3 to 10 membered cycloalkylene.
  • L 1 may be substituted or unsubstituted 3 to 6 membered cycloalkylene.
  • L 1 may be substituted 3 to 6 membered cycloalkylene.
  • L 1 may be unsubstituted 3 to 6 membered cycloalkylene.
  • L 1 may be substituted or unsubstituted 3 membered cycloalkylene.
  • L 1 may be substituted or unsubstituted 4 membered cycloalkylene.
  • L 1 may be substituted or unsubstituted 5 membered cycloalkylene.
  • L 1 may be substituted or unsubstituted 6 membered cycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted cycloalkylene.
  • L 1 may be R 14 -substituted cycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 3 to 10 membered cycloalkylene.
  • L 1 may be R 14 -substituted 3 to 10 membered cycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 3 to 6 membered cycloalkylene.
  • L 1 may be R 14 -substituted 3 to 6 membered cycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 3 membered cycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 4 membered cycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 5 membered cycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 6 membered cycloalkylene.
  • L 1 may be substituted or unsubstituted heterocycloalkylene.
  • L 1 may be substituted heterocycloalkylene.
  • L 1 may be unsubstituted heterocycloalkylene.
  • L 1 may be substituted or unsubstituted 3 to 10 membered heterocycloalkylene.
  • L 1 may be substituted 3 to 10 membered heterocycloalkylene.
  • L 1 may be unsubstituted 3 to 10 membered heterocycloalkylene.
  • L 1 may be substituted or unsubstituted 3 to 6 membered heterocycloalkylene.
  • L 1 may be substituted 3 to 6 membered heterocycloalkylene.
  • L 1 may be unsubstituted 3 to 6 membered heterocycloalkylene.
  • L 1 may be substituted or unsubstituted 3 membered heterocycloalkylene. L 1 may be substituted or unsubstituted 4 membered heterocycloalkylene. L 1 may be substituted or unsubstituted 5 membered heterocycloalkylene. L 1 may be substituted or unsubstituted 6 membered heterocycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted heterocycloalkylene.
  • L 1 may be R 14 -substituted heterocycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 3 to 10 membered heterocycloalkylene.
  • L 1 may be R 14 -substituted 3 to 10 membered heterocycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 3 to 6 membered heterocycloalkylene.
  • L 1 may be R 14 -substituted 3 to 6 membered heterocycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 3 membered heterocycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 4 membered heterocycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 5 membered heterocycloalkylene.
  • L 1 may be R 14 -substituted or unsubstituted 6 membered heterocycloalkylene.
  • L 1 may be substituted or unsubstituted arylene.
  • L 1 may be substituted arylene.
  • L 1 may be unsubstituted arylene.
  • L 1 may be substituted or unsubstituted 5 to 10 membered arylene.
  • L 1 may be substituted 5 to 10 membered arylene.
  • L 1 may be unsubstituted 5 to 10 membered arylene.
  • L 1 may be substituted or unsubstituted 5 membered arylene.
  • L 1 may be substituted 5 membered arylene.
  • L 1 may be unsubstituted 5 membered arylene.
  • L 1 may be substituted or unsubstituted 6 membered arylene.
  • L 1 may be substituted 6 membered arylene.
  • L 1 may be unsubstituted 6 membered arylene (e.g. phenylene).
  • L 1 may be R 14 -substituted or unsubstituted arylene.
  • L 1 may be R 14 -substituted arylene.
  • L 1 may be R 14 -substituted or unsubstituted 5 to 10 membered arylene.
  • L 1 may be R 14 -substituted 5 to 10 membered arylene.
  • L 1 may be R 14 -substituted or unsubstituted 5 membered arylene.
  • L 1 may be R 14 -substituted 5 membered arylene.
  • L 1 may be R 14 -substituted or unsubstituted 6 membered arylene.
  • L 1 may be R 14 -substituted 6 membered arylene.
  • L 1 may be substituted or unsubstituted heteroarylene.
  • L 1 may be substituted heteroarylene.
  • L 1 may be unsubstituted heteroarylene.
  • L 1 may be substituted or unsubstituted 5 to 10 membered heteroarylene.
  • L 1 may be substituted 5 to 10 membered heteroarylene.
  • L 1 may be unsubstituted 5 to 10 membered heteroarylene.
  • L 1 may be substituted or unsubstituted 5 membered heteroarylene.
  • L 1 may be substituted 5 membered heteroarylene.
  • L 1 may be unsubstituted 5 membered heteroarylene.
  • L 1 may be substituted or unsubstituted 6 membered heteroarylene.
  • L 1 may be substituted 6 membered heteroarylene.
  • L 1 may be unsubstituted 6 membered heteroarylene.
  • L 1 may be R 14 -substituted or unsubstituted heteroarylene.
  • L 1 may be R 14 -substituted heteroarylene.
  • L 1 may be R 14 -substituted or unsubstituted 5 to 10 membered heteroarylene.
  • L 1 may be R 14 -substituted 5 to 10 membered heteroarylene.
  • L 1 may be R 14 -substituted or unsubstituted 5 membered heteroarylene.
  • L 1 may be R 14 -substituted 5 membered heteroarylene.
  • L 1 may be R 14 -substituted or unsubstituted 6 membered heteroarylene.
  • L 1 may be R 14 -substituted 6 membered heteroarylene.
  • R 14 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , R 15 -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 15 -substituted or unsubstituted (e.g.
  • R 15 -substituted or unsubstituted e.g. C 3 -C 8 cycloalkyl
  • R 15 -substituted or unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl e.g. 1,3-butaned
  • R s1 -substituted or unsubstituted aryl e.g. phenyl or naphthyl
  • R 15 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 14 may be hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, substituted or unsubstituted (e.g.
  • heteroalkyl substituted or unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • substituted or unsubstituted e.g. C 3 -C 8
  • substituted or unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl substituted or unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 14 may be hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , unsubstituted (e.g. C 1 -C 5 ) alkyl, unsubstituted (e.g.
  • heteroalkyl 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • unsubstituted e.g. C 3 -C 8
  • unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • R 15 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , R 16 -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 16 -substituted or unsubstituted (e.g.
  • R 16 -substituted or unsubstituted e.g. C 3 -C 8 cycloalkyl
  • R 16 -substituted or unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl e.g. 1,3-butaned
  • R 16 -substituted or unsubstituted aryl e.g. phenyl or naphthyl
  • R 16 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 16 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , R 17 -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 17 -substituted or unsubstituted (e.g.
  • R 17 -substituted or unsubstituted e.g. C 3 -C 8 cycloalkyl
  • R 17 -substituted or unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl e.g. 1,3-butaned
  • R 17 -substituted or unsubstituted aryl e.g. phenyl or naphthyl
  • R 17 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 17 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , unsubstituted (e.g. C 1 -C 5 ) alkyl, unsubstituted (e.g.
  • heteroalkyl 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • unsubstituted e.g. C 3 -C 8
  • unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • R 6 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 6A , —C(O)R 6A , —NR 6A R 6B , —C(O)OR 6A , —C(O)NR 6A R 6B , —NO 2 , —SR 6A , —S(O) n6 R 6A , —S(O) n6 OR 6A , —S(O) n6 NR 6A R 6B , —NHNR 6A R 6B , —ONR 6A R 6B , or —NHC(O)NHNR 6A R 6B .
  • R 6 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 6A , —C(O)R 6A , —NR 6A R 6B , —C(O)OR 6A , —C(O)NR 6A R 6B , —NO 2 , —SR 6A , or —S(O) n6 R 6A .
  • R 6 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 6A , —NR 6A R 6B , —C(O)OR 6A , —C(O)NR 6A R 6B , —NO 2 , or —SR 6A .
  • R 6 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 6A , —NR 6A R 6B , —C(O)OR 6A , —NO 2 , or —SR 6A .
  • R 6 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 6A , —NR 6A R 6B , or —NO 2 .
  • R 6 may be hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 6A , or —NH 2 .
  • R 6 may be hydrogen.
  • R 6 may be halogen.
  • R 6 may be —N 3 .
  • R 6 may be —CF 3 .
  • R 6 may be —CN.
  • R 6 may be —OR 6A .
  • R 6 may be —NH 2 .
  • R 6 may be —NO 2 .
  • R 6 may be substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • R 6 may be substituted or unsubstituted alkyl.
  • R 6 may be substituted alkyl.
  • R 6 may be unsubstituted alkyl.
  • R 6 may be substituted or unsubstituted C 1 -C 20 alkyl.
  • R 6 may be substituted C 1 -C 20 alkyl.
  • R 6 may be substituted or unsubstituted C 1 -C 10 alkyl.
  • R 6 may be substituted C 1 -C 10 alkyl.
  • R 6 may be unsubstituted C 1 -C 10 alkyl.
  • R 6 may be substituted or unsubstituted C 1 -C 5 alkyl.
  • R 6 may be substituted C 1 -C 5 alkyl.
  • R 6 may be unsubstituted C 1 -C 5 alkyl.
  • R 6 may be substituted or unsubstituted C 1 -C 3 alkyl.
  • R 6 may be substituted C 1 -C 3 alkyl.
  • R 6 may be unsubstituted C 1 -C 3 alkyl.
  • R 6 may be methyl.
  • R 6 may be ethyl.
  • R 6 may be propyl.
  • R 6 may be R 60 -substituted or unsubstituted alkyl.
  • R 6 may be R 60 -substituted alkyl.
  • R 6 may be R 60 -substituted or unsubstituted C 1 -C 20 alkyl.
  • R 6 may be R 60 -substituted C 1 -C 20 alkyl.
  • R 6 may be R 60 -substituted or unsubstituted C 1 -C 10 alkyl.
  • R 6 may be R 60 -substituted C 1 -C 10 alkyl
  • R 6 may be R 60 -substituted or unsubstituted C 1 -C 5 alkyl.
  • R 6 may be R 60 -substituted C 1 -C 5 alkyl.
  • R 6 may be R 60 -substituted or unsubstituted C 1 -C 3 alkyl.
  • R 6 may be R 60 -substituted C 1 -C 3 alkyl.
  • R 6 may be substituted or unsubstituted 2 to 20 membered heteroalkyl.
  • R 6 may be substituted 2 to 20 membered heteroalkyl.
  • R 6 may be unsubstituted 2 to 20 membered heteroalkyl.
  • R 6 may be substituted or unsubstituted 2 to 10 membered heteroalkyl.
  • R 6 may be substituted 2 to 10 membered heteroalkyl.
  • R 6 may be unsubstituted 2 to 10 membered heteroalkyl.
  • R 6 may be substituted or unsubstituted 2 to 6 membered heteroalkyl.
  • R 6 may be substituted 2 to 6 membered heteroalkyl.
  • R 6 may be substituted or unsubstituted 2 to 6 membered heteroalkyl.
  • R 6 may be substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 6 may be substituted 2 to 5 membered heteroalkyl.
  • R 6
  • R 6 may be R 60 -substituted or unsubstituted 2 to 20 membered heteroalkyl.
  • R 6 may be R 60 -substituted 2 to 20 membered heteroalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 2 to 10 membered heteroalkyl.
  • R 6 may be R 60 -substituted 2 to 10 membered heteroalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 2 to 6 membered heteroalkyl.
  • R 6 may be R 60 -substituted 2 to 6 membered heteroalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 6 may be R 60 -substituted 2 to 5 membered heteroalkyl.
  • R 6 may be substituted or unsubstituted 3 to 10 membered cycloalkyl.
  • R 6 may be substituted 3 to 10 membered cycloalkyl.
  • R 6 may be unsubstituted 3 to 10 membered cycloalkyl.
  • R 6 may be substituted or unsubstituted 3 to 6 membered cycloalkyl.
  • R 6 may be substituted 3 to 6 membered cycloalkyl.
  • R 6 may be unsubstituted 3 to 6 membered cycloalkyl.
  • R 6 may be substituted or unsubstituted 3 to 5 membered cycloalkyl.
  • R 6 may be substituted 3 to 5 membered cycloalkyl.
  • R 6 may be unsubstituted 3 to 5 membered cycloalkyl.
  • R 6 may be substituted or unsubstituted 3 membered cycloalkyl.
  • R 6 may be substituted 3 membered cycloalkyl.
  • R 6 may be unsubstituted 3 membered cycloalkyl.
  • R 6 may be substituted or unsubstituted 4 membered cycloalkyl.
  • R 6 may be substituted 4 membered cycloalkyl.
  • R 6 may be unsubstituted 4 membered cycloalkyl.
  • R 6 may be substituted or unsubstituted 5 membered cycloalkyl.
  • R 6 may be substituted 5 membered cycloalkyl.
  • R 6 may be unsubstituted 5 membered cycloalkyl.
  • R 6 may be substituted or unsubstituted 6 membered cycloalkyl.
  • R 6 may be substituted 6 membered cycloalkyl.
  • R 6 may be unsubstituted 6 membered cycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 3 to 10 membered cycloalkyl.
  • R 6 may be R 60 -substituted 3 to 10 membered cycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 3 to 6 membered cycloalkyl.
  • R 6 may be R 60 -substituted 3 to 6 membered cycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 3 to 5 membered cycloalkyl.
  • R 6 may be R 60 -substituted 3 to 5 membered cycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 3 membered cycloalkyl.
  • R 6 may be R 60 -substituted 3 membered cycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 4 membered cycloalkyl.
  • R 6 may be R 60 -substituted 4 membered cycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 5 membered cycloalkyl.
  • R 6 may be R 60 -substituted 5 membered cycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 6 membered cycloalkyl.
  • R 6 may be R 60 substituted 6 membered cycloalkyl.
  • R 6 may be substituted or unsubstituted 3 to 10 membered heterocycloalkyl.
  • R 6 may be substituted 3 to 10 membered heterocycloalkyl.
  • R 6 may be unsubstituted 3 to 10 membered heterocycloalkyl.
  • R 6 may be substituted or unsubstituted 3 to 6 membered heterocycloalkyl.
  • R 6 may be substituted 3 to 6 membered heterocycloalkyl.
  • R 6 may be unsubstituted 3 to 6 membered heterocycloalkyl.
  • R 6 may be substituted or unsubstituted 3 to 5 membered heterocycloalkyl.
  • R 6 may be substituted 3 to 5 membered heterocycloalkyl.
  • R 6 may be unsubstituted 3 to 5 membered heterocycloalkyl.
  • R 6 may be substituted or unsubstituted 3 membered heterocycloalkyl.
  • R 6 may be substituted 3 membered heterocycloalkyl.
  • R 6 may be unsubstituted 3 membered heterocycloalkyl.
  • R 6 may be substituted or unsubstituted 4 membered heterocycloalkyl.
  • R 6 may be substituted 4 membered heterocycloalkyl.
  • R 6 may be unsubstituted 4 membered heterocycloalkyl.
  • R 6 may be substituted or unsubstituted 5 membered heterocycloalkyl.
  • R 6 may be substituted 5 membered heterocycloalkyl.
  • R 6 may be unsubstituted 5 membered heterocycloalkyl.
  • R 6 may be substituted or unsubstituted 6 membered heterocycloalkyl.
  • R 6 may be substituted 6 membered heterocycloalkyl.
  • R 6 may be unsubstituted 6 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 3 to 10 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted 3 to 10 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 3 to 6 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted 3 to 6 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 3 to 5 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted 3 to 5 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 3 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted 3 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 4 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted 4 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 5 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted 5 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted or unsubstituted 6 membered heterocycloalkyl.
  • R 6 may be R 60 -substituted 6 membered heterocycloalkyl.
  • R 6 may be substituted or unsubstituted aryl.
  • R 6 may be substituted aryl.
  • R 6 may be unsubstituted aryl.
  • R 6 may be substituted or unsubstituted 5 to 10 membered aryl.
  • R 6 may be substituted 5 to 10 membered aryl.
  • R 6 may be unsubstituted 5 to 10 membered aryl.
  • R 6 may be substituted or unsubstituted 5 or 6 membered aryl.
  • R 6 may be substituted 5 or 6 membered aryl.
  • R 6 may be unsubstituted 5 or 6 membered aryl.
  • R 6 may be substituted or unsubstituted 5 membered aryl.
  • R 6 may be substituted 5 membered aryl.
  • R 6 may be unsubstituted 5 membered aryl.
  • R 6 may be substituted or unsubstituted 6 membered aryl.
  • R 6 may be substituted 6 membered aryl.
  • R 6 may be unsubstituted 6 membered aryl.
  • R 6 may be R 60 -substituted or unsubstituted aryl.
  • R 6 may be R 60 -substituted aryl.
  • R 6 may be R 60 -substituted or unsubstituted 5 to 10 membered aryl.
  • R 6 may be R 60 -substituted 5 to 10 membered aryl.
  • R 6 may be R 60 -substituted or unsubstituted 5 or 6 membered aryl.
  • R 6 may be R 60 -substituted 5 or 6 membered aryl.
  • R 6 may be R 60 -substituted or unsubstituted 5 membered aryl.
  • R 6 may be R 60 -substituted 5 membered aryl.
  • R 6 may be R 60 -substituted or unsubstituted 6 membered aryl.
  • R 6 may be R 60 -substituted 6 membered aryl.
  • R 6 may be substituted or unsubstituted heteroaryl.
  • R 6 may be substituted heteroaryl.
  • R 6 may be unsubstituted heteroaryl.
  • R 6 may be substituted or unsubstituted 5 to 10 membered heteroaryl.
  • R 6 may be substituted 5 to 10 membered heteroaryl.
  • R 6 may be unsubstituted 5 to 10 membered heteroaryl.
  • R 6 may be substituted or unsubstituted 5 or 6 membered heteroaryl.
  • R 6 may be substituted 5 or 6 membered heteroaryl.
  • R 6 may be unsubstituted 5 or 6 membered heteroaryl.
  • R 6 may be substituted or unsubstituted 5 membered heteroaryl.
  • R 6 may be substituted or unsubstituted 5 membered heteroaryl.
  • R 6 may be substituted 5 membered heteroaryl.
  • R 6 may be unsubstituted 5 membered heteroaryl.
  • R 6 may be substituted or unsubstituted 6 membered heteroaryl.
  • R 6 may be substituted 6 membered heteroaryl.
  • R 6 may be unsubstituted 6 membered heteroaryl.
  • R 6 may be R 60 -substituted or unsubstituted heteroaryl.
  • R 6 may be R 60 -substituted heteroaryl.
  • R 6 may be R 60 -substituted or unsubstituted 5 to 10 membered heteroaryl.
  • R 6 may be R 60 -substituted 5 to 10 membered heteroaryl.
  • R 6 may be R 60 -substituted or unsubstituted 5 or 6 membered heteroaryl.
  • R 6 may be R 60 -substituted 5 or 6 membered heteroaryl.
  • R 6 may be R 60 -substituted or unsubstituted 5 membered heteroaryl.
  • R 6 may be R 60 -substituted 5 membered heteroaryl.
  • R 6 may be R 60 -substituted or unsubstituted 6 membered heteroaryl.
  • R 6 may be R 60 -substituted 6 membered heteroaryl.
  • R 60 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , R 61 -substituted or unsubstituted (e.g.
  • R 61 -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl R 61 -substituted or unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, R 61 -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 61 -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 61 -substituted or unsubstituted (e.g.
  • R 60 may be hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , substituted or unsubstituted (e.g.
  • C 1 -C 5 alkyl substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, substituted or unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • substituted or unsubstituted e.g. 2 to 5 membered
  • substituted or unsubstituted e.g. C 3 -C 8
  • cycloalkyl substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl
  • substituted or unsubstituted aryl e.g. phenyl or
  • R 60 may be hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , unsubstituted (e.g. C 1 -C 5 ) alkyl, unsubstituted (e.g.
  • heteroalkyl 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • unsubstituted e.g. C 3 -C 8
  • unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • R 61 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , R 62 -substituted or unsubstituted alkyl, R 62 -substituted or unsubstituted heteroalkyl, R 62 -substituted or unsubstituted cycloalkyl, R 62 -substituted or unsubstituted heterocycloalkyl, R 62 -substituted or unsubstituted aryl, or R
  • R 62 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , R 63 -substituted or unsubstituted (e.g.
  • R 63 -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl R 63 -substituted or unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, R 63 -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 63 -substituted or unsubstituted e.g. phenyl or naphthyl), or R 63 -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • R 63 is hydrogen, halogen, oxo, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , unsubstituted (e.g. C 1 -C 5 ) alkyl, unsubstituted (e.g.
  • heteroalkyl 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • unsubstituted e.g. C 3 -C 8
  • unsubstituted e.g. 3 to 8 membered
  • heterocycloalkyl unsubstituted e.g. 3 to 8 membered
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • R 6A and R 6B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, or —ONH 2 .
  • R 6A and R 6B may independently be substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • R 6A and R 6B may independently be hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 3 H, —ONH 2 , —NHC(O)NHNH 2 , R 60 -substituted or unsubstituted (e.g. C1-C 5 ) alkyl, R 60 -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, R 60 -substituted or unsubstituted (e.g.
  • C 3 -C 8 cycloalkyl, R 60 -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 60 -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 60 -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • aryl e.g. phenyl or naphthyl
  • R 60 -substituted or unsubstituted e.g. 5 or 6 membered or fused ring
  • R 6A and R 6B may independently be R 6C -substituted or unsubstituted (e.g. C 1 -C 5 ) alkyl, R 6C -substituted or unsubstituted (e.g. 2 to 5 membered) heteroalkyl, R 6C -substituted or unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, R 6C -substituted or unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, R 6C -substituted or unsubstituted aryl (e.g. phenyl or naphthyl), or R 6C -substituted or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • R 6C -substituted or unsubstituted e.g. C 1 -
  • R 6C is hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 Cl, —S(O) 3 H, —S(O) 4 H, —S(O) 2 NH 2 , —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , —NHC(O)NH 2 , —NHS(O) 2 H, —NHC(O)H, —NHC(O)—OH, —NHOH, —OCF 3 , —OCHF 2 , unsubstituted (e.g.
  • C 1 -C 5 alkyl unsubstituted (e.g. 2 to 5 membered) heteroalkyl, unsubstituted (e.g. C 3 -C 8 ) cycloalkyl, unsubstituted (e.g. 3 to 8 membered) heterocycloalkyl, unsubstituted aryl (e.g. phenyl or naphthyl), or unsubstituted (e.g. 5 or 6 membered or fused ring) heteroaryl.
  • aryl e.g. phenyl or naphthyl
  • unsubstituted e.g. 5 or 6 membered or fused ring
  • the compound of formula (I) may have the formula:
  • R 2 of the compounds of formula (II) or (III) may be halogen or —OR 2A , where R 2A is hydrogen or unsubstituted C 1 -C 5 alkyl compounds.
  • X of formula (II) or (III) may be —CH 2 — or —N—.
  • Y of formula (II) or (III) may be —NH— or —O—.
  • L 1 of formula (II) or (III) may be a bond.
  • R 1 of formula (II) or (III) may be halogen, —NO 2 , —NH 2 , —OR 1A , where R 1A is hydrogen or unsubstituted C 1 -C 5 alkyl, where the symbol m1 is 1, 2, or 3.
  • the compound of formula (I) may have the formula:
  • R 1 may be —OR 1A , where R 1A is as described herein (e.g. substituted or unsubstituted alkyl) and m1 is 1, 2, or 3.
  • R 1 may be halogen.
  • R 1 may be —CF 3 .
  • R 1 may be —NO 2 .
  • R 1 may be —NH 2 .
  • R 1 may be substituted at the 2, 3, or 4 positions with one or more of be —OR 1A , where R 1A is as described herein (e.g. substituted or unsubstituted alkyl), halogen, —CF 3 , —NO 2 , or —NH 2 .
  • the symbol m1 may be 1, 2, or 3.
  • the compound of formula (I) may have the formula:
  • L 1 , R 1 , m1, R 2 and X are as described herein.
  • L 1 may be a bond and R 1 may be —OR 1A , where R 1A is as defined herein (e.g. substituted or unsubstituted alkyl) and m1 is 1, 2, or 3.
  • R 1 may be halogen.
  • R 1 may be —CF 3 .
  • R 1 may be —NO 2 .
  • R 1 may be —NH 2 .
  • R 1 may be substituted at the 2, 3, or 4 positions with one or more of be —OR 1A , where R 1A is as described herein (e.g. substituted or unsubstituted alkyl), halogen, —CF 3 , —NO 2 , or —NH 2 .
  • the symbol m1 may be 1, 2, or 3.
  • the compound of formula (I) may have formula:
  • R 2 of the compounds of formula (IV) or formula (V) may be halogen or —OR 2A , where R 2A is hydrogen or unsubstituted C 1 -C 5 alkyl.
  • X of the compounds of formula (IV) or formula (V) may be —CH 2 — or —N—.
  • Y of the compounds of formula (IV) or formula (V) may be —NH— or —O—.
  • L 1 of the compounds of formula (IV) or formula (V) may be a bond.
  • R 1 of the compounds of formula (IV) or formula (V) may be halogen, —NO 2 , —NH 2 , —OR 1A , where R 1A is hydrogen or unsubstituted C 1 -C 5 alkyl where the symbol m1 is 1, 2, or 3.
  • the compound of formula (I) may be a compound having the structure set forth in Table 1, 2, or 3, or in the examples provided herein.
  • R 1 and R 2 of the compound of formula (I) are not unsubstituted aryl. In embodiments R 1 and R 2 of the compound of formula (I) are not unsubstituted phenyl. In embodiments, when L 1 is a bond, R 1 is not unsubstituted aryl or unsubstituted heteroaryl. In embodiments, when L 1 is a bond, R 1 is not substituted or unsubstituted aryl (e.g. phenyl). In embodiments, when L 1 is a bond, R 1 is not —OR 1A , where R 1A is methyl or substituted or unsubstituted benzyl.
  • R 1 when L 1 is a bond, R 1 is not —OR 1A , where R 1A is methyl or substituted or unsubstituted benzyl and R 2 is not —OCH 3 .
  • R 1 when L 1 is substituted or unsubstituted arylene, R 1 is not substituted or unsubstituted aryl.
  • L 1 when L 1 is unsubstituted arylene (e.g. phenyl), R 1 is not substituted or unsubstituted aryl.
  • L 1 when L 1 is unsubstituted arylene, R 1 is not unsubstituted aryl (e.g. phenyl).
  • the compound is not a compound as set forth in Scheme 1. In embodiments the compound is not a compound as set forth in Scheme 2. In embodiments the compound is not a compound as set forth in Scheme 3. In embodiments the compound is not a compound as set forth in Scheme 4. In embodiments the compound is not a compound as set forth in Scheme 5. In embodiments the compound is not a compound as set forth in Scheme 6. In embodiments the compound is not a compound as set forth in Table 1. In embodiments the compound is not a compound as set forth in Table 4. In embodiments the compound is not a compound as set forth in Table 5. In embodiments the compound is not a compound as set forth in Table 6.
  • the compound is not a compound having formula:
  • the compound is not a compound having formula:
  • the compound is not a compound having formula:
  • the compound is not a compound having formula:
  • the compound is not a compound having formula:
  • the method is a method of treating cancer by administering to a subject in need thereof an effective amount of a HDAC8 inhibitor as described herein.
  • the HDAC8 inhibitor is as described herein.
  • the HDAC8 inhibitor may be a compound of formula (I) as described herein including one or more compounds set forth in Tables 1-3 and/or one or more of those set forth in the Examples.
  • the HDAC8 inhibitor may be an HDAC8 inhibitor antibody.
  • the HDAC8 inhibitor may be a HDAC8 inhibitor polynucleotide.
  • the HDAC8 inhibitor may be a HDAC8 inhibitor protein. The inhibitor may block the active site of HDAC8.
  • the cancer may be a non-mutated p53 cancer.
  • the non-mutated p53 cancer may be a blood cancer (e.g. a non-mutated p53 blood cancer) or a solid tumor cancer (e.g. a non-mutated p53 solid tumor cancer).
  • the non-mutated p53 cancer may be acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), lymphoma, neuroblastoma, glioma, bladder cancer, lung cancer, non-small cell lung cancer, or breast cancer (including triple-negative breast cancer).
  • the non-mutated p53 cancer may be acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), lymphoma, neuroblastoma, glioma, bladder cancer, or lung cancer.
  • the non-mutated p53 cancer may be acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), lymphoma, neuroblastoma, or glioma.
  • the non-mutated p53 cancer may be acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or lymphoma.
  • the non-mutated p53 cancer may be acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL).
  • the non-mutated p53 cancer may be acute myeloid leukemia (AML).
  • the non-mutated p53 cancer may be acute lymphoblastic leukemia (ALL).
  • the non-mutated p53 cancer may be lymphoma.
  • the non-mutated p53 cancer may be neuroblastoma.
  • the non-mutated p53 cancer may be glioma.
  • the non-mutated p53 cancer may be bladder cancer.
  • the non-mutated p53 cancer may be lung cancer.
  • the non-mutated p53 cancer may be non-small cell lung cancer.
  • the non-mutated p53 cancer may be breast cancer (including triple-negative breast cancer).
  • the cancer may have increased HDAC8 activity or expression when compared to a non-cancerous cell.
  • the cancer may have increased HDAC8 activity and expression when compared to a non-cancerous cell.
  • the cancer may have increased HDAC8 activity when compared to a non-cancerous cell.
  • the cancer may have increased HDAC8 expression when compared to a non-cancerous cell.
  • the non-mutated p53 cancer may have increased HDAC8 activity or expression when compared to a mutated p53 cancer.
  • the non-mutated p53 cancer may have increased HDAC8 activity and expression when compared to a mutated p53 cancer.
  • the non-mutated p53 cancer may have increased HDAC8 activity when compared to a mutated p53 cancer.
  • the non-mutated p53 cancer may have increased HDAC8 expression when compared to a mutated p53 cancer.
  • p53 may be deacetylated in the non-mutated cancer.
  • p53 may be deacetylated in a non-mutated cancer having increased HDAC8 activity or expression.
  • the non-mutated p53 cancer may have deacetylated p53 and may be characterized by increased resistance to treatment with an anti-cancer agent when compared to a cancer having mutated p53.
  • the non-mutated p53 cancer may have increased HDAC8 expression and may be characterized by increased resistance to treatment with an anti-cancer agent when compared to a cancer having mutated p53.
  • the methods described herein may further include determining whether the non-mutated cancer has increased HDAC8 activity or HDAC8 expression.
  • Increased HDAC8 activity or HDAC8 expression may be determined using techniques known in the art.
  • the level of HDAC8 activity may be measured from a sample taken from the subject and compared to the level of HDAC8 activity in a control sample (e.g. a healthy or non-cancerous cell).
  • the level of HDAC8 expression may be determined using methods of quantifying a polypeptide (e.g. fluorometric or colorimetric assays, IHC, ELISA, or western blots).
  • the level of HDAC8 expression may be determined using methods of quantifying a polynucleotide (e.g. fluorometric or colorimetric assays, PCR, or northern blots).
  • the method may further include determining the expression or activity levels of HDAC8 in the non-mutated p53 cancer. When a non-mutated p53 cancer in the subject is determined to also have increased activity or expression of HDAC8, the cancer may be resistant to anti-cancer agents. When the non-mutated cancer in the subject is determined to also have increased activity or expression of HDAC8, p53 in the non-mutated cancer may be deacetylated.
  • the non-mutated p53 cancer may be leukemia where the cancer also has increased HDAC expression or activity.
  • the non-mutated p53 cancer may be leukemia where the cancer also has increased HDAC expression.
  • the non-mutated p53 cancer may be leukemia where the cancer also has increased HDAC activity.
  • the method is a method of treating cancer stem cells by administering to a subject in need thereof an effective amount of a HDAC8 inhibitor as described herein.
  • the HDAC inhibitor is as described herein.
  • the HDAC8 inhibitor may inhibit the leukemia-initiating capacity (e.g. population of cancer stem cells causing relapse).
  • the cancer stem cells may be resistant to treatment with anti-cancer agents listed herein.
  • treatments with an HDAC8 inhibitor described herein may target cancer stem cells resistant to such treatments and allow efficacy of previously resistant anti-cancer agents.
  • the HDAC8 inhibitor may be selective for HDAC8 over other HDAC isoforms (e.g. HDAC1, HDAC2, HDAC3, HDAC6, HDAC10, or HDAC 11).
  • the HDAC8 inhibitor may be at least 1, 2, 5, 10, 15, 25, 50, 100, 200, 300, 400, or 500 ⁇ more selective (e.g. by determining K i or IC 50 values for the compound for HDAC8 and other HDAC isoforms described herein).
  • the HDAC8 inhibitor may be a compound described herein.
  • the cancer is as described herein.
  • the cancer may overexpress HDAC8.
  • the method includes contacting HDAC8 with a HDAC8 inhibitor as described herein, thereby inhibiting HDAC8 mediated deacetylation of p53.
  • the inhibition of HDAC8 may allow for acetylation and activation of p53, thereby mediating cell apoptosis.
  • the contacting may be performed in vitro or in vivo.
  • the contacting may be performed in vitro.
  • the contacting may be performed in vivo.
  • the contacting may be performed in an organism.
  • HDAC8 mediated deacetylation of p53 may be monitored by techniques known in the art, including for example, fluorescent and colorimetric assays.
  • the method may be a method of activating p53 in vivo by contacting a cell with a HDAC8 inhibitor in the presence of HDAC8 and allowing the HDAC8 inhibitor to contact the HDAC8, thereby inhibiting the HDAC8 and activating p53.
  • the contacting is performed as described herein.
  • the HDAC8 inhibitor is as described herein.
  • the HDAC8 inhibitor may be a compound described herein.
  • N-hydroxycinnamides 8a-f The synthesis of N-hydroxycinnamides 8a-f is illustrated in Scheme 1 following. 7-Hydroxycoumarin 3 reacted with the appropriate benzyl bromides gave corresponding coumarins 4a-f. Ethanolysis of compounds 4a-f using sodium ethoxide under anhydrous conditions provided (E)-ethyl cinnamates 5a-f,[26] respectively. Methylation of compounds 5a-f reacted with DMS gave corresponding cinnamic esters 6a-f. Saponification of compounds 6a-f in the presence of LiOH gave corresponding cinnamic acids 7a-f in quantitative yields.
  • N-Hydroxycinnamides 13a-e were achieved starting from compounds 11 a-e through saponification followed by reaction with ethyl chloroformate and hydroxylamine according to the synthetic approach for 8a-f.
  • the synthesis of N-hydroxycinnamides 18a-d is shown in Scheme 3.
  • Suzuki coupling[27] of 19 with the appropriate aryl borates using catalytic tetrakis(triphenylphosphine) palladium yielded compounds 20a-g, respectively.
  • compound 20a-g as the starting material, saponification followed by reaction with ethyl chloroformate and hydroxylamine gave corresponding the N-hydroxycinnamides 22a-g.
  • the synthesis of ortho-phenyl N-hydroxycinnamides 27a-c with various chain lengths at the para position is achieved as described in Scheme 5. Reaction of 7-hydroxycoumarin 3 with the appropriate alkyl bromides with a three- to five-carbon chain length gave 23a-c, respectively.
  • the reaction was diluted with distilled water (100 mL), acidified with 1n HCl(aq) to pH 2-3, and extracted with EtOAc (3 ⁇ 50 mL). The organic layer was dried (Na 2 SO 4 ) and filtered, and the solvent was removed in vacuo.
  • the prepared free NH 2 OH solution was then added to the reaction and stirring was continued for 3 h.
  • the reaction was diluted with distilled water (100 mL), acidified with 1n HCl(aq) to pH 2-3, and extracted with EtOAc (3_50 mL).
  • the combined organic layer was dried (Na 2 SO 4 ) and filtered, and the solvent was removed in vacuo.
  • para-benzyl N-hydroxycinnamides 8a-f and ortho-benzyl N-hydroxycinnamides 13a-e were screened for inhibitory activity against HDAC8 at a concentration of 1 ⁇ m, using SAHA as a reference compound.
  • Ortho-substituted series 13a-e showed higher potency than parasubstituted 8a-f.
  • Compounds 13a, 13c, and 13d were further evaluated for IC 50 values against HDAC8, as well as against HeLa nuclear extract that contained mainly HDACs1-3, to analyze isoform selectivity (Table 4).
  • N-hydroxycinnamides 18a-d were synthesized with various linker chains added to the ortho-aryl groups, as well as ortho-phenyl N-hydroxycinnamide 22a with a shortened linker chain (Table 5, Table 6).
  • HDAC8 inhibitors 22b, 22d, 22 f, and 22g were evaluated for anti-proliferative activity in human lung cancer cell lines, including A549 cells, H1299 cells, CL-1 cells, and CL1-5 cells using SAHA and PCI34051 as reference compounds (Table 7).
  • Compound 22b exhibited low cytotoxicity in all four cancer cell lines. In addition, compounds 22d and 22g showed higher cytotoxicity than PCI34051 in three cancer cell lines. Although compound 22d showed moderate anti-proliferative effects in human lung cancer A549 and H1299 cells, it exhibited activity similar to that of SAHA in CL1-5 cells with no significant cytotoxicity in normal IMR-90 cells. The HDAC8 level in CL1-5 is higher than that in H1299 and A549.
  • HDAC8-selective we tested inhibitory activities of compounds 22b and 22d against a panel of purified HDACs, including class I (HDAC1, 2, 3),[28] class II (HDAC4, 6, 10)[29] and class IV (HDAC11)[30] enzymes. Table 8 shows that these compounds were inactive toward most other HDACs and had limited activity against HDAC1 and 3.
  • Example 4 Inhibition of HDAC8 Reactivates p53 and Abrogates Leukemia Stem Cell Activity in CBF ⁇ -SMMHC Associated Acute Myeloid Leukemia
  • AML Acute myeloid leukemia
  • CBF core-binding factor
  • a functional CBF complex consists of a DNA-binding a subunit (RUNX1, RUNX2, RUNX3) and a non-DNA binding ⁇ subunit (CBF ⁇ ), which increases DNA-binding affinity and may be essential for transactivation activity [3-5].
  • Chromosomal 16 inversion inv(16)(p13.1q22) or t(16;16)(p13.1;q22) is found in approximately 5-12% of AML patients and is associated with the FAB M4Eo AML subtype [6-11]. This inversion results in fusion of CBFB with the MYH11 gene, which encodes a smooth muscle myosin heavy chain (SMMHC) protein [12].
  • the CBFB-MYH11 fusion gene encodes a fusion protein CBF ⁇ -SMMHC, which retains the RUNX1 binding interface of CBF ⁇ and the coiled-coil rod region of SMMHC.
  • a knock-in allele for Cbfb-MYH11 was previously generated in mice and resultant Cbfb-MYH11 heterozygotes showed a profound defect in definitive hematopoiesis and exhibited lethal hemorrhage at E12.5 [13].
  • These phenotypes are identical to those of Runx1- or Cbfb-null mice [14,15], suggesting that CBF ⁇ -SMMHC is a dominant inhibitor of CBF function.
  • CBF ⁇ -SMMHC Dominant inhibition of RUNX1 has been considered the main function of CBF ⁇ -SMMHC.
  • CBF ⁇ -SMMHC was shown to contain an additional RUNX1 binding site within SMMHC near the inversion breakpoint, and thus binds to RUNX1 with a much higher affinity than CBF ⁇ [17]. Therefore, it is proposed that CBF ⁇ -SMMHC transdominantly inhibits RUNX1 function through high affinity binding and cytoplasmic sequestration [18,19]. Meanwhile, CBF ⁇ -SMMHC was shown to interact with the mSin3A corepressor and histone deacetylase 8 (HDAC8), supporting an alternative model where CBF ⁇ -SMMHC converts RUNX1 to a constitutive transcriptional repressor [20,21].
  • HDAC8 histone deacetylase 8
  • the tumor suppressor p53 is a genomic guardian that centrally coordinates cellular responses including cell cycle progression, DNA repair, and apoptosis. Genetic mutations that inactivate p53 function occur in approximately half of all cases of human cancer; however, these TP53 mutations are relatively rare in de novo AML (approximately 10%). TP53 mutation in AML is correlated with complex karyotype, drug resistance and dismal outcome [26-28]. Loss of p53 is shown to promote AML pathogenesis in mice by enabling aberrant self-renewal [29].
  • HDACs are a family of enzymes that catalyze the removal of acetyl moieties from ⁇ -amino groups of lysine residues in a variety of proteins, including histones and transcription factors, and thus play crucial roles in chromatin remodeling and regulation of gene expression.
  • HDAC8 is a class I HDAC, maps to the X chromosome q13 [31-33], and is likely important for diverse biological functions, including smooth muscle contraction [34], telomere protection [35], skull morphogenesis [36], and regulation of cohesion dynamics [37].
  • HDAC8 is highly expressed in myeloid and lymphoid leukemia cell lines [33] and is likely overexpressed in multiple tumor types, including neuroblastoma, glioma [38] and childhood acute lymphoblastic leukemia [39]. Although HDAC8 has been shown to interact with CBF ⁇ -SMMHC [21], its role in AML pathogenesis and maintenance remains unclear.
  • mice used were backcrossed to C57BL/6 for more than 8 generations.
  • Ai14 Cre reporter tdTomato mice on C57BL/6 background were obtained from the Jackson Laboratory.
  • To induce CBF ⁇ -SMMHC expression 4-6 week old Mx1-Cre/Cbfb 56M/+ mice were injected with 250 ⁇ g of polyinosinic-polycytidylic acid (pIpC) (InvivoGen) every 2 days for 7 doses.
  • pIpC polyinosinic-polycytidylic acid
  • mice treated Cbfb 56M/+ littermates without Cre were used as control.
  • Pre-leukemic cells were isolated from mice 2 weeks after the last dose of pIpC treatment.
  • mice were monitored up to 6 months and analyzed when moribund.
  • Transplantation of AML cells (2 ⁇ 10 6 cells/mouse) was performed via tail vein injection into sub-lethally irradiated (6.5 Gy) 6-8-week-old congenic C57BL/6 mice (CD45.1 + /CD45.2 + ). All mice were maintained in an AAALAC-accredited animal facility at City of Hope, and all experimental procedures involving mice were performed in accordance with federal and state government guidelines and established institutional guidelines and protocols approved by the Institutional Animal Care and Use Committee at Beckman Research Institute of City of Hope.
  • CB samples were provided by Stemcyte (Arcadia, Calif.). Mobilized peripheral blood stem cells (PBSC) were obtained from healthy donors (City of Hope). Inv(16) + AML samples were obtained from previously untreated patients at the City of Hope. See Table 6X following for characteristics of human samples for studies disclosed herein.
  • CD34 + cell isolation was performed using magnetic beads (StemCell Technologies, Vancouver, BC, Canada).
  • Leukopheresis samples were processed for CD34 + cell selection with CliniMACS (Miltenyi Biotech, Germany). All subjects signed an informed consent form. Sample acquisition was approved by the Institutional Review Boards at the City of Hope, in accordance with an assurance filed with and approved by the Department of Health and Human Services.
  • Bone marrow cells were isolated as previously described [59]. Cell culture employed conditions well known in the art. Bone marrow cells or 32D cells were transduced with MSCV-ires-GFP (MIG) based retroviruses or lentiviruses (pLKO.1 or HIV-7) [40] by spinoculation in the presence of 5 ug/mL polybrene (American Bioanalytical, Natick, Mass.). Human CD34 + cells were transduced with pLKO.1 lentivirus by two rounds of spinoculation.
  • MIG MSCV-ires-GFP
  • pLKO.1 or HIV-7 lentiviruses
  • Human CD34 + cells were transduced with pLKO.1 lentivirus by two rounds of spinoculation.
  • Cell sorting was performed on a 4-laser, 15-detector FACSAria-III or a 6-laser, 18-detector FACSAria II SORP (BD Bioscience, San Jose, Calif.). Cell proliferation and apoptosis assays were conducted as well known in the art.
  • IP Immunoprecipitation
  • Horseradish peroxidase-conjugated anti-rabbit or anti-mouse secondary antibodies were used, followed by detection using the SuperFemto kit (Pierce Biotechnology, Rockford, Ill.).
  • the DUOLINK® kit (Sigma, St. Louis, Mo.) was used to perform in situ proximity ligation assay (PLA) following manufacturer suggested procedures.
  • Antibodies used included mouse anti-CBF ⁇ (Santa Cruz), rabbit anti-Ac-p53 (K379) (Cell Signaling) and rabbit anti-p53 (Cell Signaling). Slides were mounted using in anti-fade media containing DAPI (Santa Cruz) and imaged using a Zeiss upright LSM 510 2-photon confocal microscope.
  • CBF ⁇ -SMMHC exerts its leukemogenic function through affecting molecular and cellular processes operating to safeguard genome integrity. Since p53 represents the master genomic guardian, we tested whether CBF ⁇ -SMMHC fusion protein impairs p53 function.
  • CM CBF ⁇ -SMMHC
  • MIG MSCV-ires-GFP
  • Acetylation of p53 protein is important for protein stabilization and its transcriptional activity.
  • CM acetylated
  • K379 acetylated p53
  • the level of Ac-p53 was markedly reduced in CM-expressing cells compared to control cells expressing CBF ⁇ at all time points (2, 4, 6, 12 h) analyzed ( FIG. 1B ).
  • Cbfb 56M/+ /Mx1-Cre can be efficiently induced to express CM by pIpC [16].
  • CM-expression To rule out possible secondary pre-leukemic effects not directly caused by CM-expression, we introduced Cre through MIG retroviral transduction of BM progenitors from Cbfb 56M/+ mice. Transduced cells were sorted 48 h later and tested for IR (3 Gy)-induced p53 acetylation. CM expression was efficiently induced in MIG-Cre-transduced progenitor cells and readily led to marked reduction in the Ac-p53 level ( FIG. 1D ), suggesting this likely to be a primary effect of CM. Therefore, we tested whether knocking-down CM cells could restore p53 acetylation in CM-expressing.
  • CBF ⁇ -SMMHC Forms an Aberrant Protein Complex with p53 and HDAC8.
  • CM interacts with p53 in primary hematopoietic cells expressing CM at endogenous levels we performed similar co-IP and western blot analysis using pre-leukemic or leukemic cells isolated from pIpC induced Cbfb 56M/+ /Mx1-Cre or Cbfb 56M/+ control mice. Consistent with results from 32D cells, we found that p53 forms a protein complex with CM in pre-leukemic as well as leukemic progenitor cells with or without IR ( FIG. 2C ). To assess the cellular localization of this complex, we isolated nuclear and cytoplasmic fractions for co-IP using anti-Flag followed by western blot with anti-p53 antibodies.
  • CM fusion protein is present in both the nucleus and the cytoplasm, however, the complex with p53 is detected exclusively in the nucleus ( FIG. 2D ).
  • PKA Duolink in situ proximity ligation assay
  • Similar Duolink in situ PLA using an Ac-p53 (K379) specific antibody showed very few interacting foci ( FIG. 9 ), suggesting that CM interacts with mostly deacetylated p53 proteins.
  • CM forms a multimeric complex with HDAC8 and p53 by sequential IP followed by western blot analysis to detect co-immunoprecipitating proteins.
  • CM- ⁇ C95 deletion does not bind HDAC8 ( FIG. 10 ), however, it was capable of binding to p53 as detected by co-IP and Duolink PLA ( FIG. 3C , D).
  • CM deletion mutants lacking regions containing the high affinity RUNX binding sites [18]. Both d134 (residues 134-236 deleted) and d179 (residues 179-221 deleted) deletion mutants were unable to bind p53 ( FIG. 3C , D).
  • CM and p53 may be independent of HDAC8 binding.
  • lentivirus (pLKO.1) mediated expression of 2 independent small-hairpin (sh)-RNA sequences against Hdac8 to specifically knock-down Hdac8 in 32D-CM cells ( FIG. 3E , left).
  • the binding of CM and p53 is unaffected in Hdac8 knocked-down cells ( FIG. 3E right, F), confirming that the protein-protein interaction between CM and p53 is independent of HDAC8.
  • HDAC8 Mediates the Deacetylation of p53 Associated with CBF ⁇ -SMMHC.
  • HDACs including HDAC1, HDAC2, and HDAC3, reportedly could inhibit p53 activity by deacetylating p53 [41-43].
  • HDAC8 mediates the aberrant deacetylation of p53 in CM-expressing cells.
  • lentivirus pLKO.1 mediated expression of 2 independent small-hairpin (sh)-RNA sequences against Hdac8 to specifically knock-down Hdac8 in 32D-CM cells.
  • HDAC8i HDAC8 isoform-selective inhibitors
  • PCI-34051 [44] or compound 22d [45] directed against its catalytic activity.
  • Treatment with both HDAC8i remarkably increased Ac-p53 in CM-expressing cells ( FIG. 4B ). Since p53 protein levels were also increased upon HDAC8i treatment, we included Mdm2 inhibitor Nutlin-3 to stabilize p53 protein.
  • HDAC8i treatment PCI-34051 or 22d
  • Nutlin-3 enhanced Ac-p53 compared to Nutlin-3 alone
  • FIG. 4B HDAC8i (22d) treatment did not disrupt the interaction of CM with p53, further supporting the involvement of deacetylase activity.
  • expression of the CM- ⁇ C95 deletion mutant that was unable to bind HDAC8 had no effect on Ac-p53 induction compared to control (FLAG or CBF ⁇ ) 32D cells ( FIG. 4C ).
  • HDAC8 Activates p53 and Selectively Induces Apoptosis of Inv(16) + AML Stem and Progenitor Cells.
  • HDAC8 altered post-translational modification by HDAC8 could represent an alternative p53-inactivating mechanism and contribute to drug resistance of AML stem cells.
  • PBSC peripheral blood stem cells
  • 22d HDAC8i selectively induced apoptosis of inv(16) + AML CD34 + cells compared to normal CD34 + cells ( FIG.
  • AML LSCs are functionally characterized by their capacity to engraft and reproduce AML disease in transplanted host.
  • HDAC8i p53 activation induced by HDAC8i could reduce or eliminate LSC engraftment and leukemia-initiating capacity.
  • CM-induced AML model allowing for high-level engraftment and reproducible disease upon secondary transplantation.
  • Cbfb 56M/+ /Mx1-Cre mice with a Cre-reporter line expressing tdTomato fluorescence protein following Cre-mediated recombination.
  • AML can be induced in Cbfb 56M/+ /Mx1-Cre/tdTomato + mice 3-6 months following pIpC and the AML cells are predominately dTomato + /cKit + (data not shown).
  • Freshly isolated AML cells (2 ⁇ 10 6 ) from BM of moribund animals were treated with 22d (10 ⁇ M) or vehicle for 48 h before transplanting into sub-lethally irradiated congenic recipients ( FIG. 6A ).
  • Progression of AML disease is evident by increasing frequencies of dTomato + /cKit + cells in the peripheral blood of mice transplanted with vehicle-treated cells whereas these cells are barely detectable in mice receiving 22d-treated cells ( FIG. 6B ).
  • mice in the vehicle-treated group all showed enlarged spleens compared to the 22d-treated group ( FIGS. 6C, 15A ).
  • 2 out of 7 mice in the vehicle-treated group succumbed to lethal AML 5-6 weeks after transplantation prior to analysis of BM and spleen engraftment (data not shown).
  • AML transformation requires multiple genetic and/or epigenetic alterations, which cooperatively impair differentiation, and confer proliferation and survival signals.
  • CBF ⁇ -SMMHC expression dominantly inhibits CBF/RUNX1 function and disrupts hematopoietic differentiation [13,16,46,47]. It has been long thought that dominant inhibition of CBF/RUNX1 function underlies the leukemogenic function of CBF ⁇ -SMMHC.
  • CBF ⁇ -SMMHC-mediated leukemogenesis requires functional Runx proteins [22], and that RUNX1 dominant inhibition is not essential for transformation [23].
  • RUNX1 activity is now shown to be required for the growth and survival of AML cells, including RUNX1-ETO and CBF ⁇ -SMMHC associated leukemia [24,25].
  • CBF ⁇ -SMMHC binds to target DNA in a RUNX1-dependent manner [48].
  • CBF ⁇ -SMMHC fusion protein disrupts p53 activity through aberrant post-translational modification. It has been previously reported that CBF ⁇ -SMMHC reduces p53 mRNA transcription and slows apoptosis in Ba/F3 pro-B cell line [49].
  • Histone deacetylases including HDAC1, 2, 3 and Sirtuin I (SIRT1) are known to modulate p53 activity through deacetylating p53 [41-43,50].
  • HDAC1 HDAC1
  • SIRT1 Sirtuin I
  • CBF ⁇ -SMMHC reduced acetylated p53 levels particularly at later time points after stimulation ( FIG. 1B , C), suggesting that p53 is aberrantly deacetylated.
  • HDAC8 similar to other class I HDACs, can deacetylate p53 and modulate p53 activity.
  • AML is maintained by LSCs that are relatively resistant to chemotherapy and can persist as potential sources of relapse. Even though AML with CBF translocations are considered to have favorable prognosis, the incidence of relapse is still 25-58% with chemotherapy [51-53]. Novel strategies directed to eradicate LSCs are required to improve treatment outcome.
  • HDAC8 an isotype-selective inhibitor, 22d, leads to restoration of p53 acetylation and activity, induction of apoptosis, and abrogates engraftment and leukemia-initiating activity of inv(16) + AML LSCs. Whether additional mechanisms besides increased apoptosis contribute to the reduced engraftment capacity remains to be examined.
  • this effect is selective for LSCs as normal HSCs display relatively low levels of p53 target activation ( FIG. 5E ) and induction of apoptosis ( FIG. 5C ).
  • Treatment of 22d also had no impact on short-term or long-term engraftment activity of normal HSCs ( FIG. 18 ).
  • This selectivity is likely due to the combined effect of elevated HDAC8 expression and the recruitment of HDAC8 and p53 into a stable protein complex in inv(16) + AML CD34 + cells.
  • the increase in HDAC8 expression does not appear to be directly caused by CM expression since we did not observe changes of HDAC8 levels in 32D cells expressing CM ( FIG. 19 ).
  • the impact of CM-associated protein complex on HDAC8 activity requires further investigation.
  • HDAC8 function can potentially be exploited to modulate p53 activity in AML and other cancers overexpressing HDAC8. Indeed, inhibition of HDAC8 results in reduced clonogenic growth and enhanced differentiation in neuroblastoma where high HDAC8 expression is associated with poor prognosis [38]. Most broad-spectrum HDAC inhibitors currently used or being tested in clinical trails display low activity against HDAC8 [57].
  • Example 9 HDAC8 Inhibition Activates p53 and Induces p53-Dependent Apoptosis in Human AML Cells
  • FIG. 21B demonstrates that increased HDAC8i concentration results in greater signal for Ac-p53 and p53 by western blot analysis to assay the effects of administration of HDAC8i (e.g. 22d) for 6-hr for Ac-p53, p53, Ac-H3, Ac-H4 and ⁇ -actin.
  • HDAC8i e.g. 22d
  • FIG. 21C The fold change in mRNA levels of p53 targets after treatment with PCI-48012 for 16-hrs is depicted in FIG. 21C .
  • FIG. 21D Western blot analysis of p53, b-actin and MV4-11 cells transduced with control or sh-p53 is provided in FIG. 21D .
  • Relative survival of non-p53 mutated AML cell lines (MV4-11, MOLM13, and OCI-AML3) treated with HDAC8i compounds (Cmp[ds 22d, 5b, 5e, 5h) for 48-hr was determined by Annexin V labeling and normalization to vehicle-treated controls. See FIGS. 22A-22C .
  • Relative proliferation of AML cell lines (MV4-11 and MOLM13) treated with HDAC8i (Cmpds 22d, 5b, 5e, 5h) for 48-hr was determined by luminescent cell viability assay, normalized to vehicle treated control. See FIGS. 22D-22E .
  • HDAC8i compounds Resulting inhibitory activity of HDAC8i compounds is tabulated in Table 14 following.
  • CBF ⁇ -SMMHC reduces p53 mRNA transcription and slows apoptosis in Ba/F3 pro-B cell line.
  • FIG. 26A Expression of after contact with Cmpd 22d on Ac-p53, p53 and ⁇ -actin under various timing and wash conditions are depicted in FIG. 26B .
  • FIG. 26C The survival rate with and without wash of Cmpd 22d is depicted in FIG. 26C , showing that washing of Cmpd 22d results in increased survival.
  • Embodiment P1 A compound having formula:
  • R 1 is substituted or unsubstituted C 1 -C 20 alkyl, substituted or unsubstituted 2 to 20 membered heteroalkyl, substituted or unsubstituted 3 to 7 membered cycloalkyl, substituted or unsubstituted 3 to 7 membered heterocycloalkyl, substituted or unsubstituted 3 to 7 membered aryl, or substituted or unsubstituted 3 to 7 membered heteroaryl; and R 2 is substituted or unsubstituted C 1 -C 20 alkyl, substituted or unsubstituted 2 to 20 membered heteroalkyl, substituted or unsubstituted 3 to 7 membered cycloalkyl, substituted or unsubstituted 3 to 7 membered heterocycloalkyl, substituted or unsubstituted 3 to 7 membered aryl, or substituted or unsubstituted 3 to 7 membered heteroaryl.
  • Embodiment P2 The compound of embodiment P1 having the formula:
  • Embodiment P3 The compound of any one of embodiments P1 to P2, wherein said compound is a HDAC8 inhibitor.
  • Embodiment P4 A method of treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount the compound of any one of embodiments P1 to P3.
  • Embodiment P5 The method of embodiment P4, wherein said cancer is a hematological cancer.
  • Embodiment P6 The method of any one of embodiments P4 to P5, wherein said cancer is acute myeloid leukemia.
  • Embodiment 1 A compound having the formula:
  • A is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.
  • X is —C(R 4 ) or —N ⁇ .
  • Y is a bond, —N(R 5 )—, —O—, or —S—.
  • L 1 is a bond, —C(O)—, —C(O)O—, —O—, —S—, —N(R 6 )—, —C(O)N(R 6 )—, —S(O) 6 —, —S(O)N(R 6 )—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene.
  • R 1 is halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 1A , —C(O)R 1A , —NR 1A R 1B , —C(O)OR 1A , —C(O)NR 1A R 1B , —NO 2 , —SR 1A , —S(O) n1 R 1A , —S(O) n1 OR 1A , —S(O) n1 NR 1A R 1B , —NHNR 1A R 1B , —ONR 1A R 1B , —NHC(O)NHNR 1A R 1B , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substitute
  • R 2 is halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 2A , —C(O)R 2A , —NR 2A R 2B , —C(O)OR 2A , —C(O)NR 2A R 2B , —NO 2 , —SR 2A , —S(O) n2 R 2A , —S(O) n2 OR 2A , —S(O) n2 NR 2A R 2B , —NHNR 2A R 2B , —ONR 2A R 2B , —NHC(O)NHNR 2A R 2B , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 3 is independently hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 H, —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 4 is hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 4A , —C(O)R 4A , —NR 4A R 4B , —C(O)OR 4A , —C(O)NR 4A R 4B , —NO 2 , —SR 4A , —S(O) n4 R 4A , —S(O) n4 OR 4A , —S(O) n4 NR 4A R 4B , —NHNR 4A R 4B , —ONR 4A R 4B , —NHC(O)NHNR 4A R 4B , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 5 is hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 5A , —C(O)R 5A , —NR 5A R 5B , —C(O)OR 5A , —C(O)NR 5A R 5B , —NO 2 , —SR 5A , —S(O) n5 R 5A , —S(O) n5 OR 5A , —S(O) n5 NR 5A R 5B , —NHNR 5A R 5B , —ONR 5A R 5B , —NHC(O)NHNR 5A R 5B , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • R 6 is hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —OR 6A , —C(O)R 6A , —NR 6A R 6B , —C(O)OR 6A , —C(O)NR 6A R 6B , —NO 2 , —SR 6A , —S(O) n6 R 6A , —S(O) n6 OR 6A , —S(O)NR 6A R 6B , —NHNR 6A R 6B , —ONR 6A R 6B , —NHC(O)NHNR 6A R 6B , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or
  • R 1A , R 1B , R 2A , R 2B , R 4A , R 4B , R 5A , R 5B , R 6A , and R 6B are independently hydrogen, oxo, halogen, —CF 3 , —CN, —OH, —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O) 2 Cl, —S(O) 3 H, —S(O) 4 H, —S(O) 2 NH 2 , —NHNH 2 , —ONH 2 , —NHC(O)NHNH 2 , —NHC(O)NH 2 , —NHS(O) 2 H, —NHC(O)H, —NHC(O)—OH, —NHOH, —OCF 3 , —OCHF 2 , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl
  • Embodiment 2 The compound of embodiment 1, wherein X is —C(R 4 )—.
  • Embodiment 3 The compound of any one of embodiments 1 to 2, wherein R 2 is halogen, —CF 3 , —OR 2A , —NO 2 , substituted or unsubstituted C 1 -C 5 alkyl, or substituted or unsubstituted 2 to 5 membered heteroalkyl.
  • Embodiment 4 The compound of one of embodiments 1 to 3, wherein R 2 is OR 2A , wherein R 2A is substituted or unsubstituted C 1 -C 3 alkyl.
  • Embodiment 5 The compound of one of embodiments 1 to 4, wherein R 3 is hydrogen, halogen, or —OR 3A .
  • Embodiment 6 The compound of embodiment 5, wherein R 3 is hydrogen.
  • Embodiment 7 The compound of one of embodiments 1 to 6, wherein m2 is 0.
  • Embodiment 8 The compound of one of embodiments 1 to 7, wherein Y is a bond or —N(R 5 )—.
  • Embodiment 9 The compound of embodiment 8, wherein Y is a bond or —NH—.
  • Embodiment 10 The compound of one of embodiments 1 to 9, wherein L 1 is a bond, —C(O)—, —O—, —NH—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene.
  • Embodiment 11 The compound of one of embodiments 1 to 10, wherein R 1 is halogen, —CF 3 , —NO 2 , —NH 2 , —OR 1A , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
  • Embodiment 12 The compound of one of embodiments 1 to 11, wherein R 1 is halogen, —CF 3 , —NO 2 , —NH 2 , —OR 1A .
  • Embodiment 13 The compound of one of embodiments 1 to 12, wherein R 1 is —OR 1A , wherein R 1A is substituted or unsubstituted C 1 -C 5 alkyl, substituted or unsubstituted 2 to 5 membered heteroalkyl, substituted or unsubstituted 3 to 6 membered cycloalkyl, substituted or unsubstituted 3 to 6 membered heterocycloalkyl, substituted or unsubstituted 5 or 6 membered aryl, or substituted or unsubstituted 5 or 6 membered heteroaryl.
  • Embodiment 14 The compound of one of embodiments 1 to 13, wherein R 1 is R 10 -substituted or unsubstituted alkyl, R 10 -substituted or unsubstituted heteroalkyl, R 10 -substituted or unsubstituted cycloalkyl, R 10 -substituted or unsubstituted heterocycloalkyl, R 10 -substituted or unsubstituted aryl, or R 10 -substituted or unsubstituted heteroaryl, wherein R 10 is hydrogen, halogen, —N 3 , —CF 3 , —CCl 3 , —CBr 3 , —CI 3 , —CN, —C(O)H, —OCH 3 , —OCH 2 CH 3 , —NH 2 , —COOH, —CONH 2 , —NO 2 , —SH, —S(O)
  • Embodiment 15 The compound of one of embodiments 1 to 14, wherein m1 is 1, 2, or 3.
  • Embodiment 16 The compound of one of embodiments 1 to 15, wherein A is aryl or heteroaryl.
  • Embodiment 17 The compound of one of embodiments 1 to 16, wherein said compound has the formula:
  • Embodiment 18 The compound of embodiment 17, wherein
  • Embodiment 19 The compound of one of embodiments 1 to 16, wherein A is aryl, 5,6-fused ring heteroaryl, 6,5-fused ring heteroaryl, or 6,6-fused ring heteroaryl.
  • Embodiment 20 The compound of embodiment 19, wherein said compound has the formula:
  • A is 5,6-fused ring heteroaryl, 6,5-fused ring heteroaryl, or 6,6-fused ring heteroaryl.
  • Embodiment 21 The compound of embodiment 20, wherein
  • Embodiment 22 A method of treating cancer in a subject in need thereof, said method comprising administering an effective amount of an HDAC8 inhibitor to said subject.
  • Embodiment 23 The method of embodiment 22, wherein said cancer is a non-mutated p53 cancer.
  • Embodiment 24 The method of any one of embodiments 22 to 23, wherein said non-mutated p53 cancer is acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), lymphoma, neuroblastoma, glioma, bladder cancer, lung cancer, non-small cell lung cancer, breast cancer, or triple-negative breast cancer.
  • AML acute myeloid leukemia
  • ALL acute lymphoblastic leukemia
  • lymphoma lymphoma
  • neuroblastoma neuroblastoma
  • glioma bladder cancer
  • lung cancer non-small cell lung cancer
  • breast cancer breast cancer
  • triple-negative breast cancer triple-negative breast cancer
  • Embodiment 25 The method of any one of embodiments 22 to 24, wherein said non-mutated p53 cancer is acute myeloid leukemia (AML).
  • AML acute myeloid leukemia
  • Embodiment 26 The method of any one of embodiments 22 to 25, wherein said non-mutated p53 cancer has increased HDAC8 activity or HDAC8 expression compared to a mutated p53 cancer.
  • Embodiment 27 The method of any one of embodiments 22 to 26, wherein said method further comprises prior to said treating, determining whether said cancer in said subject is a non-mutated p53 cancer.
  • Embodiment 28 The method of any one of embodiments 22 to 27, further comprising determining whether said non-mutated p53 cancer has increased HDAC8 activity or HDAC8 expression.
  • Embodiment 29 The method of any one of embodiments 22 to 28, wherein said HDAC8 inhibitor is a HDAC8 inhibitor compound.
  • Embodiment 30 The method of any one of embodiments 22 to 29, wherein said HDAC8 inhibitor is a compound described herein.
  • Embodiment 31 The method of any one of embodiments 22 to 28, wherein said HDAC8 inhibitor is a HDAC8 inhibitor antibody, a HDAC8 inhibitor siRNA, a HDAC8 inhibitor shRNA, or a HDAC8 inhibitor protein.
  • said HDAC8 inhibitor is a HDAC8 inhibitor antibody, a HDAC8 inhibitor siRNA, a HDAC8 inhibitor shRNA, or a HDAC8 inhibitor protein.
  • Embodiment 32 A method of inhibiting HDAC8 mediated deacetylation of p53, said method comprising contacting HDAC8 with a HDAC8 inhibitor in the presence of p53, thereby inhibiting HDAC8 deacetylation of p53.
  • Embodiment 33 The method of embodiment 31, wherein said contacting is performed in vitro.
  • Embodiment 34 The method of embodiment 31, wherein said contacting is performed in vivo.
  • Embodiment 35 The method of embodiment 31, wherein said contacting is performed in an organism.
  • Embodiment 36 The method of any one of embodiments 32 to 35, wherein said HDAC8 inhibitor is a compound described herein.
  • Embodiment 37 A method of activating p53 in vivo, said method comprising contacting a cell with a HDAC8 inhibitor in the presence of HDAC8 and allowing said HDAC8 inhibitor to contact said HDAC8, thereby inhibiting said HDAC8 and activating p53.
  • Embodiment 38 The method of embodiment 36, wherein said contacting is performed in an organism.
  • Embodiment 39 The method of any one of embodiments 37 to 38, wherein said HDAC8 inhibitor is a compound described herein.

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US10465115B2 (en) * 2015-12-17 2019-11-05 Merck Patent Gmbh Cinnamic acid derivatives
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Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4861760A (en) 1985-10-03 1989-08-29 Merck & Co., Inc. Ophthalmological composition of the type which undergoes liquid-gel phase transition
US4911920A (en) 1986-07-30 1990-03-27 Alcon Laboratories, Inc. Sustained release, comfort formulation for glaucoma therapy
US5028629A (en) * 1990-03-28 1991-07-02 Eli Lilly And Company 5-Lipoxygenase inhibitors
US5212162A (en) 1991-03-27 1993-05-18 Alcon Laboratories, Inc. Use of combinations gelling polysaccharides and finely divided drug carrier substrates in topical ophthalmic compositions
US5403841A (en) 1991-01-15 1995-04-04 Alcon Laboratories, Inc. Use of carrageenans in topical ophthalmic compositions
WO2001042437A2 (en) 1999-12-08 2001-06-14 Axys Pharmaceuticals, Inc. Histone deacetylase-8 proteins, nucleic acids, and methods of use
WO2008101186A1 (en) 2007-02-15 2008-08-21 The J. David Gladstone Institutes Inhibitors for hdac8
US20120322769A1 (en) 2010-02-16 2012-12-20 Duke University Methods of reducing virulence in bacteria
WO2013101600A1 (en) 2011-12-29 2013-07-04 Pharmacyclics, Inc. Cinnamic acid hydroxyamides as inhibitors of histone deacetylase 8

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006016680A1 (en) * 2004-08-09 2006-02-16 Astellas Pharma Inc. Hydroxyamide compounds having activity as inhibitors of histone deacetylase (hdac)
US7994357B2 (en) * 2009-04-03 2011-08-09 Naturewise Biotech & Medicals Corporation Cinamic compounds and derivatives therefrom for the inhibition of histone deacetylase
JP2016530273A (ja) * 2013-08-20 2016-09-29 シティ・オブ・ホープＣｉｔｙ ｏｆ Ｈｏｐｅ 癌を治療するためのｈｄａｃ８阻害剤

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4861760A (en) 1985-10-03 1989-08-29 Merck & Co., Inc. Ophthalmological composition of the type which undergoes liquid-gel phase transition
US4911920A (en) 1986-07-30 1990-03-27 Alcon Laboratories, Inc. Sustained release, comfort formulation for glaucoma therapy
US5028629A (en) * 1990-03-28 1991-07-02 Eli Lilly And Company 5-Lipoxygenase inhibitors
US5403841A (en) 1991-01-15 1995-04-04 Alcon Laboratories, Inc. Use of carrageenans in topical ophthalmic compositions
US5212162A (en) 1991-03-27 1993-05-18 Alcon Laboratories, Inc. Use of combinations gelling polysaccharides and finely divided drug carrier substrates in topical ophthalmic compositions
WO2001042437A2 (en) 1999-12-08 2001-06-14 Axys Pharmaceuticals, Inc. Histone deacetylase-8 proteins, nucleic acids, and methods of use
WO2008101186A1 (en) 2007-02-15 2008-08-21 The J. David Gladstone Institutes Inhibitors for hdac8
US20120322769A1 (en) 2010-02-16 2012-12-20 Duke University Methods of reducing virulence in bacteria
WO2013101600A1 (en) 2011-12-29 2013-07-04 Pharmacyclics, Inc. Cinnamic acid hydroxyamides as inhibitors of histone deacetylase 8

Non-Patent Citations (72)

* Cited by examiner, † Cited by third party
Title
Adya, N. et al. (Dec. 1998). "The leukemic protein core binding factor beta (CBFbeta)-smooth-muscle myosin heavy chain sequesters CBFalpha2 into cytoskeletal filaments and aggregates," Mol Cell Biol 18(12):7432-7443.
Appelbaum, F.R., et al. (Oct. 2006). "The clinical spectrum of adult acute myeloid leukaemia associated with core binding factor translocations," Br J Haematol 135(2):165-173.
Balasubramanian, S.et al. (May 2008, e-published Feb. 7, 2008). "A novel histone deacetylase 8 (HDAC8)-specific inhibitor PCI-34051 induces apoptosis in T-cell lymphomas," Leukemia 22(5):1026-1034.
Ben-Ami, O., et al. (Sep. 26, 2013, e-published Sep. 19, 2013). "Addiction of t(8;21) and inv(16) acute myeloid leukemia to native RUNX1," Cell Rep 4(6):1131-1143.
Britos-Bray, M., et al. (Dec. 1998). "CBFbeta-SMMHC, expressed in M4eo acute myeloid leukemia, reduces p53 induction and slows apoptosis in hematopoietic cells exposed to DNA-damaging agents" Blood 92(11):4344-4352.
Buggy, J.J. et al. (Aug. 15, 2000). "Cloning and characterization of a novel human histone deacetylase, HDAC8," Biochem J 350 Pt 1:199-205.
Castilla, L.H. et al. (Nov. 15, 1996). "Failure of embryonic hematopoiesis and lethal hemorrhages in mouse embryos heterozygous for a knocked-in leukemia gene CBFB-MYH11," Cell 87(4):687-696.
Dai, C. et al. (Nov. 2010). "p53 post-translational modification: deregulated in tumorigenesis ," Trends Mol Med 16(11):528-536.
Deardorff, M.A., et al. (Sep. 13, 2012). "HDAC8 mutations in Cornelia de Lange syndrome affect the cohesin acetylation cycle," Nature 489(7415):313-317.
Durst, K.L. et al. (Jan. 2003). "The inv(16) fusion protein associates with corepressors via a smooth muscle myosin heavy-chain domain," Mol Cell Biol 23(2):607-619.
European Search Report dated Mar. 17, 2017, for EP Application No. 14837390.5, Aug. 20, 2014, 10 pages.
Galletti, P. et al. (Dec. 2009). "Azetidinones as zinc-binding groups to design selective HDAC8 inhibitors," ChemMedChem 4(12):1991-2001.
Goyama, S. et al. (Sep. 2013). "Transcription factor RUNX1 promotes survival of acute myeloid leukemia cells," J Clin Invest 123(9):3876-3888.
Grimwade, D. et al. (2009). "Independent prognostic factors for AML outcome," Hematology Am Soc Hematol Educ Program 385-395.
Grimwade, D. et al. (Jul. 22, 2010, e-published Apr. 12, 2010). "Refinement of cytogenetic classification in acute myeloid leukemia: determination of prognostic significance of rare recurring chromosomal abnormalities among 5876 younger adult patients treated in the United Kingdom Medical Research Council trials," Blood 116(3):354-365.
Haberland, M. et al. (Jul. 15, 2009). "Epigenetic control of skull morphogenesis by histone deacetylase 8," Genes Dev 23(14):1625-1630.
Haferlach, C. et al. (Aug. 2008). "Mutations of the TP53 gene in acute myeloid leukemia are strongly associated with a complex aberrant karyotype," Leukemia 22(8):1539-1541.
Hu, E. et al. (May 19, 2000). "Cloning and characterization of a novel human class I histone deacetylase that functions as a transcription repressor," J Biol Chem 275(20):15254-15264.
Hu, E. et al. (Nov. 2003, e-published Sep. 15, 2003). "Identification of novel isoform-selective inhibitors within class | histone deacetylases," J Pharmacol Exp Ther 307(2):720-728.
Huang, G. et al. (Feb. 15 2001). "Dimerization with PEBP2beta protects RUNX1/AML1 from ubiquitin-proteasome-mediated degradation," Embo J 20(4):723-733.
Huang, W.J. et al. (Oct. 2012, e-published Aug. 20, 2012). "Synthesis and biological evaluation of ortho-aryl N-hydroxycinnamides as potent histone deacetylase (HDAC) 8 isoform-selective inhibitors," ChemMedChem 7(10):1815-1824.
International Search Report dated Jan. 21, 2015, for PCT Application No. PCT/US2014/051876, filed Aug. 20, 2014, 5 pages.
Ito, A. et al. (Nov. 15, 2002). "MDM2-HDAC1-mediated deacetylation of p53 is required for its degradation," EMBO J 21(22):6236-6245.
Jeannet, R., et al. (Mar. 2013). "Alcam Regulates Long-term Hematopoietic Stem Cell Engraftment and Self-Renewal," Stem Cells 31(3): 26 pages.
Juan, L.J. et al. (Jul. 7, 2000). "Histone deacetylases specifically down-regulate p53-dependent gene activation," J Biol Chem 275(27):20436-20443.
Kamikubo, Y. et al. (May 18, 2010). "Accelerated leukemogenesis by truncated CBF beta-SMMHC defective in high-affinity binding with RUNX1," Cancer Cell 17(5):455-468.
Kanno, Y. et al. (Jul. 1998). "Cytoplasmic sequestration of the polyomavirus enhancer binding protein 2 (PEBP2)/core binding factor alpha (CBFalpha) subunit by the leukemia-related PEBP2/CBFbeta-SMMHC fusion protein inhibits PEBP2/CBF-mediated transactivation," Mol Cell Biol 18(7):4252-4261.
Khan, N. et al. (Jan. 15, 2008). "Determination of the class and isoform selectivity of small-molecule histone deacetylase inhibitors," Biochem J 409(2):581-589.
Kojima, K. et al. (Nov. 1, 2005). "MDM2 antagonists induce p53-dependent apoptosis in AML: implications for leukemia therapy," Blood 106(9):3150-3159.
Krennhrubec, K. et al. (May 15, 2007, e-published Feb. 25, 2007). "Design and evaluation of 'Linkerless' hydroxamic acids as selective HDAC8 inhibitors," Bioorg Med Chem Lett 17(10):2874-2878.
Krennhrubec, K. et al. (May 15, 2007, e-published Feb. 25, 2007). "Design and evaluation of ‘Linkerless’ hydroxamic acids as selective HDAC8 inhibitors," Bioorg Med Chem Lett 17(10):2874-2878.
Kühn, R. et al. (Sep. 8, 1995). "Inducible gene targeting in mice," Science 269(5229):1427-1429.
Kuo, Y.-H. et al. (Feb. 1, 2008). "Cbfbeta-SMMHC impairs differentiation of common lymphoid progenitors and reveals an essential role for RUNX in early B-cell development," Blood 111(3):1543-1551.
Kuo, Y.-H. et al. (Jan. 2006). "Cbf beta-SMMHC induces distinct abnormal myeloid progenitors able to develop acute myeloid leukemia," Cancer Cell 9(1):57-68.
Kuo, Y.-H., et al. (Apr. 2, 2009, e-published Jan. 28, 2009). "Runx2 induces acute myeloid leukemia in cooperation with Cbfbeta-SMMHC in mice," Blood 113(14):3323-3332.
Kurosawa, S. et al. (Oct. 2013, e-published May 28, 2013). "Prognosis of patients with core binding factor acute myeloid leukemia after first relapse," Haematologic 98(10):1525-1531.
Lee, H., et al. (Jul. 2006). "Histone deacetylase 8 safeguards the human ever-shorter telomere 1B (hEST1B) protein from ubiquitin-mediated degradation, " Mol Cell Biol 26(14):5259-5269.
Li, L. et al. (Feb. 14, 2012). "Activation of p53 by SIRT1 inhibition enhances elimination of CML leukemia stem cell in combination with imatinib," Cancer Cell 21(2):266-281.
Liu, P. et al. (Aug. 20, 1993), "Fusion between transcription factor CBF β/PEBP2β and a myosin heavy chain in acute myeloid leukemia," Science 261(5124):1041-1044.
Liu, P.P. et al. (May 1, 1995). "Molecular pathogenesis of the chromosome 16 inversion in the M4Eo subtype of acute myeloid leukemia," Blood 85(9):2289-2302.
Lo, H.-L. et al. (Nov. 2007, e-published Sep. 6, 2007). "Inhibition of HIV-1 replication with designed miRNAs expressed from RNA polymerase II promoters," Gene Ther 14(21)1 503-1512.
Long, J. et al. (Jul. 8, 2010, e-published Apr. 19, 2010). "Multiple distinct molecular mechanisms influence sensitivity and resistance to MDM2 inhibitors in adult acute myelogenous leukemia," Blood 116(1):71-80.
Look, A.T. (Nov. 7, 1997). "Oncogenic transcription factors in the human acute leukemias," Science 278(5340):1059-1064.
Lukasik, S.M. et al. (Sep. 2002). "Altered affinity of CBFβ-SMMHC for Runx1 explains its role in leukemogenesis," Nat Struct Biol 9(9):674-679.
Luo, J. et al. (Nov. 16, 2000). "Deacetylation of p53 modulates its effect on cell growth and apoptosis," Nature 408(6810):377-381.
Luo, J. et al. (Oct. 19, 2001). "Negative control of p53 by Sir2alpha promotes cell survival under stress," Cell 107(2):137-148.
Lutterbach, B. et al. (Oct. 26, 1999). "The inv(16) encodes an acute myeloid leukemia 1 transcriptional corepressor," Proc Natl Acad Sci U S A 96(22):12822-12827.
Mandoli, A. et al. (Apr. 2014, e-published Sep. 4, 2013). "CBFB-MYH11/RUNX1 together with a compendium of hematopoietic regulators, chromatin modifiers and basal transcription factors occupies self-renewal genes in inv(16) acute myeloid leukemia," 28(4):77-778.
Moreno, D.A. et al. (Sep. 2010). "Differential expression of HDAC3, HDAC7 and HDAC9 is associated with prognosis and survival in childhood acute lymphoblastic leukaemia," Br J Haematol 150(6):665-673.
Mrózek, K. et al. (Dec. 20, 2012). "Prognostic significance of the European LeukemiaNet standardized system for reporting cytogenetic and molecular alterations in adults with acute myeloid leukemia," J Clin Oncol 30(36):4515-4523.
Mrózek, K. et al. (Jun. 2004). Cytogenetics in acute leukemia, Blood Rev 18(2):115-136.
Nahi, H et al. (Mar. 2008). "Chromosomal aberrations in 17p predict in vitro drug resistance and short overall survival in acute myeloid leukemia," Leuk Lymphoma 49(3):508-516.
Oehme, I. et al. (Jan. 1, 2009). "Histone deacetylase 8 in neuroblastoma tumorigenesis," Clin Cancer Res 15(1):91-99.
Oehme, I. et al. (Nov. 2009). "Targeting of HDAC8 and investigational inhibitors in neuroblastoma," Expert Opin Investig Drugs 18(11)1605-1617.
Ogawa, E. et al. (May 1993). "Molecular cloning and characterization of PEBP2 beta, the heterodimeric partner of a novel Drosophila runt-related DNA binding protein PEBP2 alpha," Virology 194(1):314-331.
Okuda, T. et al. (Jan. 26, 1996). "AML1, the target of multiple chromosomal translocations in human leukemia, is essential for normal fetal liver hematopoiesis," Cell 84(2):321-330.
Prébet, T. et al. (Oct. 1, 2009, e-published Aug. 31, 2009). "Acute myeloid leukemia with translocation (8;21) or inversion (16) in elderly patients treated with conventional chemotherapy: a collaborative study of the French CBF-AML intergroup," J Clin Oncol 27(28):4747-4753.
PUBCHEM CID 11687004, create date: Oct. 26, 2006, located at <https://pubchem.ncbi.nlm.nih.gov/compound/11687004> last visited Jun. 20, 2017, 13 pages.
Rücker, F.G. et al. (Mar. 1, 2012, e-published Dec. 20, 2011). "TP53 alterations in acute myeloid leukemia with complex karyotype correlate with specific copy number alterations, monosomal karyotype, and dismal outcome," Blood 119(9):2114-2121.
Speck, N.A. (Jul. 2002). "Core-binding factors in haematopoiesis and leukaemia," Nat Rev Cancer 2(7):502-513.
Suzuki, T. et al. (Mar. 2014, e-published Jan. 8, 2014). "Design, synthesis, and biological activity of NCC149 derivatives as histone deacetylase 8-selective inhibitors," ChemMedChem 9(3):657-664.
Tang, W. et al. (May 2011, e-published Feb. 2, 2011). "Discovery of histone deacetylase 8 selective inhibitors," Bioorg Med Chem Lett 21(9):2601-2605.
Van Den Wyngaert, I., et al. (Jul. 28, 2000). "Cloning and characterization of human histone deacetylase 8," FEBS Lett 478(1-2):77-83.
Von Neuhoff, C., et al. (Jun. 1, 2010). "Prognostic impact of specific chromosomal aberrations in a large group of pediatric patients with acute myeloid leukemia treated uniformly according to trial AML-BFM 98," J Clin Oncol 28(16):2682-2689.
Waltregny, D., et al. (Jun. 19, 2005). "Histone deacetylase HDAC8 associates with.smooth muscle α-actin and is essential for smooth muscle cell contractility," FASEB J 19(8):966-968.
Wang, Q., et al. (Nov. 15, 1996). "The CBFbeta subunit is essential for CBFalpha2 (AML1) function in vivo," Cell 87(4):697-708.
Wang, S. et al. (Jun. 1993). "Cloning and characterization of subunits of the T-cell receptor and murine leukemia virus enhancer core-binding factor," Mol Cell Biol 13(6):3324-3339.
Whitehead, L. et al. (Aug. 1, 2011, e-published Jun. 15, 2011). "Human HDAC isoform selectivity achieved via exploitation of the acetate release channel with structurally unique small molecule inhibitors," Bioorg Med Chem 19(15):4626-4634.
Written Opinion dated Jan. 21, 2015, for PCT Application No. PCT/US2014/051876, filed Aug. 20, 2014, 5 pages.
Yan, W. et al. (Jan. 31, 2013, e-published Mar. 5, 2012). "Histone deacetylase inhibitors suppress mutant p53 transcription via histone deacetylase 8," Oncogene 32(5):599-609.
Zhao, L., et al. (Apr. 15, 2007, e-published Dec. 21, 2006). "CBFB-MYH11 hinders early T-cell development and induces massive cell death in the thymus," Blood 109(8):3432-3440.
Zhao, Z., et al. (Jul. 1, 2010). "p53 loss promotes acute myeloid leukemia by enabling aberrant self-renewal," Genes Dev 24(13):1389-1402.

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11505523B2 (en) 2013-08-20 2022-11-22 City Of Hope HDAC8 inhibitors for treating cancer

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